%PDF-1.6
%
1 0 obj
<>
endobj
2 0 obj
<>stream
Yohan Seo, Kunhi Ryu, Jinhong Park, Dong-kyu Jeon, Sungwoo Jo, Ho K. Lee, Wan Namkung
Inhibition of ANO1 by luteolin and its cytotoxicity in human prostate cancer PC-3 cells
www.plosone.org
www.plosone.org
endstream
endobj
3 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 10 0 R/Annots 11 0 R/Contents 12 0 R/TrimBox[0 0 612 792]>>
endobj
11 0 obj
[13 0 R 14 0 R 15 0 R 16 0 R 17 0 R 18 0 R 19 0 R 20 0 R 21 0 R 22 0 R 23 0 R 24 0 R]
endobj
13 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref011)>>
endobj
14 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref013)>>
endobj
15 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref006)>>
endobj
16 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref006)>>
endobj
17 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref013)>>
endobj
18 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref017)>>
endobj
19 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref018)>>
endobj
20 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref019)>>
endobj
21 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref020)>>
endobj
22 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref006)>>
endobj
23 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref016)>>
endobj
24 0 obj
<>/Border[0 0 0]/A 25 0 R>>
endobj
25 0 obj
<>
endobj
12 0 obj
[26 0 R 27 0 R 28 0 R 29 0 R 30 0 R 31 0 R 32 0 R 33 0 R 34 0 R 35 0 R 36 0 R 37 0 R]
endobj
26 0 obj
<>stream
q
0.83 0.64 0.02 0 k
300.6992 693.2976 m
310.1669 693.2976 l
h
f*
315.3827 693.2976 m
324.8504 693.2976 l
h
f*
354.5008 654.3496 m
359.263 654.3496 l
h
f*
411.137 628.3276 m
415.8992 628.3276 l
h
f*
374.8535 563.3008 m
384.378 563.3008 l
h
f*
389.537 563.3008 m
399.0614 563.3008 l
h
f*
245.8772 537.3354 m
255.4016 537.3354 l
h
f*
319.6346 472.3087 m
329.1591 472.3087 l
h
f*
329.4992 446.3433 m
339.0236 446.3433 l
h
f*
266.1732 381.3165 m
270.9354 381.3165 l
h
f*
459.2126 120.926 m
468.737 120.926 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(inhibited)Tj
3.8437 0 Td
(cancer)Tj
2.8346 0 Td
(cell)Tj
1.5477 0 Td
(proliferation,)Tj
5.4822 0 Td
(migration)Tj
4.1896 0 Td
(and)Tj
1.7178 0 Td
(invasion,)Tj
3.8267 0 Td
(even)Tj
2.058 0 Td
(though)Tj
3.067 0 Td
(the)Tj
1.457 0 Td
(underlying)Tj
4.5921 0 Td
(mecha-)Tj
-34.6164 -1.3039 Td
(nisms)Tj
2.5738 0 Td
(are)Tj
1.4513 0 Td
(still)Tj
1.6384 0 Td
(uncertain)Tj
4.0649 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(13)Tj
0 g
(].)Tj
3.5433 0 Td
(For)Tj
1.6271 0 Td
(instance,)Tj
3.7473 0 Td
(molecular)Tj
4.218 0 Td
(and)Tj
1.7348 0 Td
(electrophysiological)Tj
8.1353 0 Td
(studies)Tj
-32.7342 -1.2982 Td
(showed)Tj
3.2484 0 Td
(strong)Tj
2.7723 0 Td
(and)Tj
1.7291 0 Td
(functional)Tj
4.3087 0 Td
(expression)Tj
4.4673 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.863 0 Td
(in)Tj
1.0375 0 Td
(human)Tj
3.0727 0 Td
(metastatic)Tj
4.2803 0 Td
(prostate)Tj
3.4299 0 Td
(cancer)Tj
2.846 0 Td
(PC-3)Tj
-35.07 -1.2983 Td
(cells,)Tj
2.1429 0 Td
(and)Tj
1.7292 0 Td
(downregulation)Tj
6.5763 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.863 0 Td
(expression)Tj
4.4673 0 Td
(by)Tj
1.1792 0 Td
(shRNA)Tj
3.1521 0 Td
(induced)Tj
3.4696 0 Td
(significant)Tj
4.388 0 Td
(reduction)Tj
4.1102 0 Td
(of)Tj
1.0148 0 Td
(cell)Tj
-36.1074 -1.2983 Td
(proliferation,)Tj
5.4935 0 Td
(metastasis)Tj
4.2746 0 Td
(and)Tj
1.7291 0 Td
(invasion)Tj
3.6113 0 Td
([)Tj
0.83 0.64 0.02 0 k
(6)Tj
0 g
(].)Tj
1.5987 0 Td
(In)Tj
1.1112 0 Td
(an)Tj
1.2019 0 Td
(orthotopic)Tj
4.4447 0 Td
(xenograft)Tj
4.0195 0 Td
(mouse)Tj
2.8573 0 Td
(model)Tj
2.7439 0 Td
(of)Tj
1.0148 0 Td
(prostate)Tj
-34.1005 -1.3039 Td
(cancer)Tj
2.8402 0 Td
(using)Tj
2.3925 0 Td
(PC-3)Tj
2.2677 0 Td
(cells,)Tj
2.1486 0 Td
(downregulation)Tj
6.5764 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.8629 0 Td
(expression)Tj
4.4617 0 Td
(by)Tj
1.1849 0 Td
(intratumoral)Tj
5.3404 0 Td
(injection)Tj
3.7588 0 Td
(of)Tj
-34.8489 -1.2982 Td
(ANO1)Tj
2.8629 0 Td
(shRNA)Tj
3.1465 0 Td
(significantly)Tj
5.091 0 Td
(inhibited)Tj
3.855 0 Td
(tumor)Tj
2.744 0 Td
(growth)Tj
3.0727 0 Td
([)Tj
0.83 0.64 0.02 0 k
(6)Tj
0 g
(].)Tj
-19.5759 -1.2983 Td
(These)Tj
2.5681 0 Td
(results)Tj
2.812 0 Td
(indicate)Tj
3.4072 0 Td
(that)Tj
1.7802 0 Td
(development)Tj
5.4084 0 Td
(of)Tj
1.0148 0 Td
(potent)Tj
2.812 0 Td
(and)Tj
1.7348 0 Td
(selective)Tj
3.5376 0 Td
(small-molecule)Tj
6.2985 0 Td
(inhibitors)Tj
4.1386 0 Td
(of)Tj
-36.7084 -1.3039 Td
(ANO1)Tj
2.8629 0 Td
(may)Tj
1.9276 0 Td
(have)Tj
2.0579 0 Td
(a)Tj
0.6576 0 Td
(therapeutic)Tj
4.7282 0 Td
(potential)Tj
3.7644 0 Td
(for)Tj
1.3833 0 Td
(treatment)Tj
4.1385 0 Td
(of)Tj
1.0148 0 Td
(prostate)Tj
3.4299 0 Td
(cancer)Tj
2.8403 0 Td
(or)Tj
1.0999 0 Td
(other)Tj
2.347 0 Td
(cancers)Tj
3.2088 0 Td
(with)Tj
-35.4611 -1.2983 Td
(high)Tj
2.0069 0 Td
(levels)Tj
2.3754 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.863 0 Td
(expression.)Tj
4.6884 0 Td
(To)Tj
1.338 0 Td
(date,)Tj
2.1316 0 Td
(few)Tj
1.6158 0 Td
(ANO1)Tj
2.8629 0 Td
(inhibitors)Tj
4.1386 0 Td
(have)Tj
2.0636 0 Td
(been)Tj
2.1203 0 Td
(identified,)Tj
4.286 0 Td
(such)Tj
2.0579 0 Td
(as)Tj
-35.5632 -1.2983 Td
(CaCC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 224.2204 575.7165 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(inh)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 234.3118 577.4173 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-A01)Tj
2.2053 0 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 269.8015 575.7165 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 279.0992 577.4173 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~1)Tj
/F11 1 Tf
1.2755 0 Td
[()]TJ
/F7 1 Tf
(M\),)Tj
2.1827 0 Td
(tannic)Tj
2.7439 0 Td
(acid)Tj
1.8709 0 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 373.2661 575.7165 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 382.5637 577.4173 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~6)Tj
/F11 1 Tf
1.2755 0 Td
[()]TJ
/F7 1 Tf
(M\),)Tj
2.1884 0 Td
(T16A)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 439.6535 575.7165 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(inh)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 449.6881 577.4173 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-A01)Tj
2.2053 0 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 485.1779 575.7165 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 494.4755 577.4173 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~1)Tj
/F11 1 Tf
1.2755 0 Td
[()]TJ
/F7 1 Tf
(M\),)Tj
2.1884 0 Td
(MONNA)Tj
-32.91 -1.3039 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 213.4488 562.7338 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 222.7464 564.3779 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~1)Tj
/F11 1 Tf
1.2755 0 Td
[()]TJ
/F7 1 Tf
(M\))Tj
1.9616 0 Td
(and)Tj
1.7291 0 Td
(idebenone)Tj
4.3937 0 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 329.7826 562.7338 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 339.137 564.3779 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~9)Tj
/F11 1 Tf
1.2755 0 Td
[()]TJ
/F7 1 Tf
(M\))Tj
1.9559 0 Td
([)Tj
0.83 0.64 0.02 0 k
(13)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(],)Tj
3.549 0 Td
(and)Tj
1.7291 0 Td
(more)Tj
2.3414 0 Td
(recently)Tj
3.4072 0 Td
(we)Tj
1.3266 0 Td
(identified)Tj
4.0592 0 Td
(a)Tj
0.6577 0 Td
(novel)Tj
-34.2139 -1.2982 Td
(small-molecule)Tj
6.2929 0 Td
(inhibitor)Tj
3.7814 0 Td
(of)Tj
1.0148 0 Td
(ANO1,)Tj
3.0896 0 Td
(Ani9)Tj
2.194 0 Td
(\(IC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 377.1779 549.7511 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(50)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 386.4755 551.3952 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(~77)Tj
1.7517 0 Td
(nM\),)Tj
2.2281 0 Td
(showing)Tj
3.5829 0 Td
(high)Tj
2.007 0 Td
(potency)Tj
3.3902 0 Td
(and)Tj
1.7291 0 Td
(selectivity)Tj
-33.3351 -1.2982 Td
(for)Tj
1.3833 0 Td
(ANO1)Tj
2.8629 0 Td
([)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(].)Tj
2.0807 0 Td
(However,)Tj
4.0762 0 Td
(the)Tj
1.4683 0 Td
(mechanisms)Tj
5.2044 0 Td
(of)Tj
1.0148 0 Td
(action)Tj
2.6984 0 Td
(and)Tj
1.7292 0 Td
(pharmacological)Tj
6.8427 0 Td
(properties)Tj
4.2803 0 Td
(of)Tj
1.0148 0 Td
(these)Tj
-34.656 -1.3039 Td
(inhibitors)Tj
4.1385 0 Td
(remain)Tj
3.0841 0 Td
(unclear,)Tj
3.4242 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
1.4683 0 Td
(ANO1)Tj
2.863 0 Td
(inhibitors)Tj
4.1441 0 Td
(are)Tj
1.4457 0 Td
(still)Tj
1.6384 0 Td
(in)Tj
1.0374 0 Td
(the)Tj
1.4684 0 Td
(early)Tj
2.1486 0 Td
(phases)Tj
2.846 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(drug)Tj
-33.919 -1.2983 Td
(discovery.)Tj
1.1962 -1.2983 Td
(Natural)Tj
3.2711 0 Td
(products)Tj
3.7531 0 Td
(have)Tj
2.0579 0 Td
(historically)Tj
4.6261 0 Td
(been)Tj
2.1203 0 Td
(a)Tj
0.6577 0 Td
(productive)Tj
4.5353 0 Td
(source)Tj
2.8346 0 Td
(of)Tj
1.0148 0 Td
(pharmaceutical)Tj
6.3666 0 Td
(leads)Tj
2.2167 0 Td
(and)Tj
-34.6504 -1.2982 Td
(therapeutic)Tj
4.7338 0 Td
(drugs,)Tj
2.6929 0 Td
(and)Tj
1.7348 0 Td
(natural)Tj
3.0784 0 Td
(products)Tj
3.753 0 Td
(and)Tj
1.7292 0 Td
(their)Tj
2.1088 0 Td
(derivatives)Tj
4.5014 0 Td
(have)Tj
2.0636 0 Td
(provided)Tj
3.8154 0 Td
(a)Tj
0.652 0 Td
(number)Tj
3.4186 0 Td
(of)Tj
1.0148 0 Td
(can-)Tj
-35.2967 -1.3039 Td
(cer)Tj
1.4343 0 Td
(chemotherapeutic)Tj
7.4381 0 Td
(agents)Tj
2.7495 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.075 0 Td
(For)Tj
1.6271 0 Td
(example,)Tj
3.7756 0 Td
(natural)Tj
3.0784 0 Td
(compounds)Tj
4.9606 0 Td
(have)Tj
2.0579 0 Td
(played)Tj
2.8233 0 Td
(an)Tj
1.2019 0 Td
(essential)Tj
-33.2217 -1.2983 Td
(role)Tj
1.7688 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.474 0 Td
(development)Tj
5.4085 0 Td
(of)Tj
1.0148 0 Td
(clinically)Tj
3.7757 0 Td
(useful)Tj
2.5965 0 Td
(anticancer)Tj
4.3879 0 Td
(agents,)Tj
2.9764 0 Td
(such)Tj
2.0636 0 Td
(as)Tj
1.0148 0 Td
(vinblastine,)Tj
4.8018 0 Td
(vincristine,)Tj
-32.3146 -1.2983 Td
(topotecan,)Tj
4.3993 0 Td
(irinotecan)Tj
4.3086 0 Td
(and)Tj
1.7348 0 Td
(taxol)Tj
2.1714 0 Td
([)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(],)Tj
2.0749 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(high)Tj
2.0068 0 Td
(structural)Tj
4.0819 0 Td
(diversity)Tj
3.6453 0 Td
(and)Tj
1.7291 0 Td
(biological)Tj
4.0932 0 Td
(efficacy)Tj
3.2201 0 Td
(of)Tj
-36.6629 -1.3039 Td
(natural)Tj
3.0727 0 Td
(products)Tj
3.7417 0 Td
(make)Tj
2.3868 0 Td
(them)Tj
2.279 0 Td
(attractive)Tj
3.9175 0 Td
(sources)Tj
3.1861 0 Td
(of)Tj
1.009 0 Td
(novel)Tj
2.3868 0 Td
(scaffolds)Tj
3.651 0 Td
(and)Tj
1.7234 0 Td
(drug)Tj
2.1033 0 Td
(leads)Tj
2.1997 0 Td
(for)Tj
1.3776 0 Td
(several)Tj
2.931 0 Td
(bio-)Tj
-35.9656 -1.2983 Td
(logical)Tj
2.8062 0 Td
(targets.)Tj
-1.61 -1.2982 Td
(Prostate)Tj
3.4752 0 Td
(cancer)Tj
2.846 0 Td
(is)Tj
0.8447 0 Td
(the)Tj
1.4684 0 Td
(most)Tj
2.211 0 Td
(common)Tj
3.8437 0 Td
(type)Tj
1.9106 0 Td
(of)Tj
1.0147 0 Td
(cancer)Tj
2.8459 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4741 0 Td
(second)Tj
3.0103 0 Td
(leading)Tj
3.1295 0 Td
(cause)Tj
2.3924 0 Td
(of)Tj
1.0148 0 Td
(cancer)Tj
-34.4066 -1.3039 Td
(deaths)Tj
2.7892 0 Td
(in)Tj
1.0375 0 Td
(men,)Tj
2.2394 0 Td
(and)Tj
1.7291 0 Td
(recent)Tj
2.7099 0 Td
(studies)Tj
2.982 0 Td
(suggest)Tj
3.1238 0 Td
(ANO1)Tj
2.8629 0 Td
(may)Tj
1.9275 0 Td
(be)Tj
1.1509 0 Td
(a)Tj
0.6519 0 Td
(promising)Tj
4.3484 0 Td
(therapeutic)Tj
4.7281 0 Td
(target)Tj
2.5115 0 Td
(for)Tj
-34.7921 -1.2983 Td
(prostate)Tj
3.4299 0 Td
(cancer)Tj
2.8459 0 Td
([)Tj
0.83 0.64 0.02 0 k
(6)Tj
0 g
(].)Tj
1.5988 0 Td
(In)Tj
1.1111 0 Td
(the)Tj
1.4684 0 Td
(present)Tj
3.1634 0 Td
(study,)Tj
2.6135 0 Td
(we)Tj
1.3267 0 Td
(performed)Tj
4.4729 0 Td
(a)Tj
0.652 0 Td
(cell-based)Tj
4.1612 0 Td
(screening)Tj
4.0535 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6576 0 Td
(collection)Tj
-33.5505 -1.2983 Td
(of)Tj
1.0091 0 Td
(natural)Tj
3.0727 0 Td
(products,)Tj
3.9742 0 Td
(a)Tj
0.6463 0 Td
(good)Tj
2.2223 0 Td
(source)Tj
2.829 0 Td
(of)Tj
1.0034 0 Td
(novel)Tj
2.3981 0 Td
(scaffolds)Tj
3.6509 0 Td
(and)Tj
1.7235 0 Td
(drug)Tj
2.1033 0 Td
(leads)Tj
2.2053 0 Td
(for)Tj
1.3833 0 Td
(target)Tj
2.5058 0 Td
(proteins,)Tj
3.7304 0 Td
(to)Tj
1.0205 0 Td
(iden-)Tj
-35.4781 -1.2982 Td
(tify)Tj
1.525 0 Td
(novel)Tj
2.4038 0 Td
(ANO1)Tj
2.8629 0 Td
(inhibitors.)Tj
4.3654 0 Td
(Screening)Tj
4.1669 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(natural)Tj
3.084 0 Td
(product)Tj
3.3902 0 Td
(library)Tj
2.8743 0 Td
(revealed)Tj
3.5206 0 Td
(that)Tj
1.7802 0 Td
(luteolin)Tj
3.2825 0 Td
(is)Tj
0.8503 0 Td
(a)Tj
-36.5892 -1.3039 Td
(potent)Tj
2.8119 0 Td
(inhibitor)Tj
3.7814 0 Td
(of)Tj
1.0148 0 Td
(ANO1.)Tj
3.0898 0 Td
(We)Tj
1.6044 0 Td
(further)Tj
3.033 0 Td
(investigated)Tj
4.9888 0 Td
(the)Tj
1.4684 0 Td
(effects)Tj
2.7269 0 Td
(of)Tj
1.0148 0 Td
(luteolin)Tj
3.2825 0 Td
(on)Tj
1.2756 0 Td
(cell)Tj
1.559 0 Td
(proliferation)Tj
-31.6513 -1.2983 Td
(and)Tj
1.7291 0 Td
(migration)Tj
4.2009 0 Td
(of)Tj
1.0148 0 Td
(PC-3)Tj
2.2677 0 Td
(prostate)Tj
3.4299 0 Td
(cancer)Tj
2.8403 0 Td
(cells)Tj
1.9219 0 Td
(expressing)Tj
4.4276 0 Td
(high)Tj
2.0069 0 Td
(levels)Tj
2.3754 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.863 0 Td
(endogenously.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 304.2141 cm
BT
/F2 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Materials)Tj
4.5071 0 Td
(and)Tj
1.989 0 Td
(methods)Tj
/F0 1 Tf
-6.4961 -1.5023 Td
(Materials)Tj
4.2283 0 Td
(and)Tj
1.8615 0 Td
(solutions)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 269.1779 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Luteolin)Tj
3.5773 0 Td
(was)Tj
1.6951 0 Td
(purchased)Tj
4.3369 0 Td
(from)Tj
2.2054 0 Td
(Santa)Tj
2.4094 0 Td
(Cruz)Tj
2.194 0 Td
(Biotechnology)Tj
5.9754 0 Td
(\(Santa)Tj
2.7553 0 Td
(Cruz,)Tj
2.4207 0 Td
(CA\))Tj
1.9049 0 Td
(and)Tj
1.7291 0 Td
(kaempferol)Tj
4.7452 0 Td
(was)Tj
-35.9487 -1.2982 Td
(purchased)Tj
4.3313 0 Td
(from)Tj
2.2053 0 Td
(Tocris)Tj
2.7609 0 Td
(Bioscience)Tj
4.4447 0 Td
(\(Ellisville,)Tj
4.1159 0 Td
(MO\).)Tj
2.4264 0 Td
(Chrysin,)Tj
3.634 0 Td
(Apigenin,)Tj
4.1669 0 Td
(Galangin,)Tj
4.1046 0 Td
(and)Tj
1.7348 0 Td
(other)Tj
-33.9248 -1.2983 Td
(chemicals,)Tj
4.3653 0 Td
(unless)Tj
2.6759 0 Td
(otherwise)Tj
4.0762 0 Td
(indicated,)Tj
4.1555 0 Td
(were)Tj
2.1203 0 Td
(purchased)Tj
4.3257 0 Td
(from)Tj
2.1996 0 Td
(Sigma)Tj
2.6703 0 Td
(\(St.)Tj
1.5533 0 Td
(Louis,)Tj
2.6419 0 Td
(MO\).)Tj
2.4151 0 Td
(The)Tj
1.7745 0 Td
(collec-)Tj
-34.9736 -1.3039 Td
(tion)Tj
1.8425 0 Td
(of)Tj
1.0148 0 Td
(bioactive)Tj
3.7984 0 Td
(natural)Tj
3.0784 0 Td
(products)Tj
3.7587 0 Td
(used)Tj
2.0579 0 Td
(for)Tj
1.3833 0 Td
(screening)Tj
4.0535 0 Td
(was)Tj
1.7008 0 Td
(prepared)Tj
3.8041 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.474 0 Td
(Spectrum)Tj
4.0762 0 Td
(collec-)Tj
-34.2479 -1.2983 Td
(tion)Tj
1.8425 0 Td
(of)Tj
1.0148 0 Td
(MicroSource)Tj
5.4084 0 Td
(Discovery)Tj
4.218 0 Td
(Systems,)Tj
3.6396 0 Td
(Inc.)Tj
1.7575 0 Td
(\(Gaylordsville,)Tj
6.0377 0 Td
(CT\).)Tj
2.0636 0 Td
(HCO3)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 486.3685 221.6692 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 488.9763 217.1905 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-buffered)Tj
3.9287 0 Td
(solution)Tj
-32.8248 -1.2982 Td
(containing)Tj
4.524 0 Td
(\(in)Tj
1.3776 0 Td
(mM\):)Tj
2.5002 0 Td
(120)Tj
1.6441 0 Td
(NaCl,)Tj
2.5285 0 Td
(5)Tj
0.6916 0 Td
(KCl,)Tj
2.0296 0 Td
(1)Tj
0.6973 0 Td
(MgCl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 382.6204 202.507 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 386.1921 204.2078 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4422 0 Td
(1)Tj
0.6973 0 Td
(CaCl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 417.6566 202.507 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 421.2283 204.2078 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4422 0 Td
(10)Tj
1.1735 0 Td
(D-glucose,)Tj
4.4844 0 Td
(2.5)Tj
1.3946 0 Td
(HEPES,)Tj
3.3902 0 Td
(and)Tj
1.7291 0 Td
(25)Tj
-34.7354 -1.2982 Td
(NaHCO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 233.4614 189.5244 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 239.1874 191.2251 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\(pH)Tj
1.8425 0 Td
(7.4\).)Tj
1.9615 0 Td
(The)Tj
1.7802 0 Td
(half-Cl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 322.5826 195.6472 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 327.3448 191.2251 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(solution)Tj
3.4809 0 Td
(was)Tj
1.6951 0 Td
(prepared)Tj
3.8041 0 Td
(by)Tj
1.1792 0 Td
(replacing)Tj
3.9117 0 Td
(65)Tj
1.1679 0 Td
(mM)Tj
1.9276 0 Td
(NaCl)Tj
2.296 0 Td
(with)Tj
1.9956 0 Td
(equimo-)Tj
-34.1912 -1.3039 Td
(lar)Tj
1.2699 0 Td
(Na-gluconate)Tj
5.6126 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(HCO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 315.6661 176.5417 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
0.4762 0.9185 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 321.8456 178.1858 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-buffered)Tj
3.9287 0 Td
(solution.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 151.9937 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cell)Tj
1.9134 0 Td
(culture)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 134.9858 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Fisher)Tj
2.6985 0 Td
(rat)Tj
1.321 0 Td
(thyroid)Tj
3.1747 0 Td
(\(FRT\))Tj
2.6589 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.126 0 Td
(stably)Tj
2.5171 0 Td
(transfected)Tj
4.6205 0 Td
(with)Tj
1.9955 0 Td
(human)Tj
3.0784 0 Td
(ANO1)Tj
2.863 0 Td
(\(abc\))Tj
2.262 0 Td
(or)Tj
1.0999 0 Td
(mouse)Tj
-32.3374 -1.2982 Td
(ANO2)Tj
2.8629 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(halide)Tj
2.6475 0 Td
(sensor)Tj
2.7893 0 Td
(YFP)Tj
1.9389 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(in)Tj
1.0375 0 Td
(previous)Tj
3.651 0 Td
(study)Tj
2.3867 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(].)Tj
2.075 0 Td
(FRT)Tj
1.9842 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(cultured)Tj
-33.681 -1.2983 Td
(in)Tj
1.0318 0 Td
(Coon`s)Tj
3.033 0 Td
(modified)Tj
3.8494 0 Td
(F12)Tj
1.7008 0 Td
(medium)Tj
3.6057 0 Td
(with)Tj
1.9955 0 Td
(10%)Tj
1.9276 0 Td
(FBS,)Tj
2.0296 0 Td
(2)Tj
0.6973 0 Td
(mM)Tj
1.9275 0 Td
(L-glutamine,)Tj
5.3688 0 Td
(100)Tj
1.6441 0 Td
(units/ml)Tj
3.617 0 Td
(penicillin)Tj
-32.4281 -1.3039 Td
(and)Tj
1.7291 0 Td
(100)Tj
/F11 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
(g/ml)Tj
2.5908 0 Td
(streptomycin)Tj
5.5105 0 Td
(at)Tj
0.9525 0 Td
(37C)Tj
2.228 0 Td
(and)Tj
1.7291 0 Td
(5%)Tj
1.4513 0 Td
(CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 392.485 94.337 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 396.0566 95.9811 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(.)Tj
0.4422 0 Td
(PC-3)Tj
2.2677 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(grown)Tj
2.8006 0 Td
(in)Tj
1.0319 0 Td
(RPMI)Tj
2.6362 0 Td
(1640)Tj
-32.8249 -1.2982 Td
(medium)Tj
3.6056 0 Td
(supplemented)Tj
5.8507 0 Td
(with)Tj
2.0012 0 Td
(10%)Tj
1.9276 0 Td
(FBS,)Tj
2.0296 0 Td
(100)Tj
1.6441 0 Td
(units/ml)Tj
3.6168 0 Td
(penicillin)Tj
3.9799 0 Td
(and)Tj
1.7291 0 Td
(100)Tj
/F11 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
(g/ml)Tj
2.5908 0 Td
(streptomycin)Tj
5.5105 0 Td
(at)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 453.2598 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Downre)Tj
3.3803 0 Td
(gulation)Tj
3.6496 0 Td
(of)Tj
1.0418 0 Td
(ANO1)Tj
2.9126 0 Td
(by)Tj
1.2543 0 Td
(luteolin)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01749)Tj
4.089 0 Td
(35)Tj
0 g
2.0906 0 Td
(March)Tj
2.9622 0 Td
(31,)Tj
1.5874 0 Td
(2017)Tj
34.3562 0 Td
(2)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
27 0 obj
<>stream
endstream
endobj
28 0 obj
<>stream
endstream
endobj
29 0 obj
<>stream
endstream
endobj
30 0 obj
<>stream
endstream
endobj
31 0 obj
<>stream
endstream
endobj
32 0 obj
<>stream
endstream
endobj
33 0 obj
<>stream
endstream
endobj
34 0 obj
<>stream
endstream
endobj
35 0 obj
<>stream
endstream
endobj
36 0 obj
<>stream
endstream
endobj
37 0 obj
<>stream
BT
66.402 0 Td
(13)Tj
ET
Q
endstream
endobj
38 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 10 0 R/Annots 40 0 R/Contents 41 0 R/TrimBox[0 0 612 792]>>
endobj
40 0 obj
[42 0 R 43 0 R]
endobj
42 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref016)>>
endobj
43 0 obj
<>/Border[0 0 0]/A 44 0 R>>
endobj
44 0 obj
<>
endobj
41 0 obj
[45 0 R 46 0 R 47 0 R 48 0 R 49 0 R 50 0 R]
endobj
45 0 obj
<>stream
q
0.83 0.64 0.02 0 k
442.715 556.7244 m
452.2394 556.7244 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(37C)Tj
2.228 0 Td
(and)Tj
1.7291 0 Td
(5%)Tj
1.4513 0 Td
(CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 268.2141 705.7133 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 271.7291 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(.)Tj
0.4478 0 Td
(There)Tj
2.5795 0 Td
(were)Tj
2.1203 0 Td
(populations)Tj
4.9493 0 Td
(of)Tj
1.0148 0 Td
(PC-3)Tj
2.2734 0 Td
(cells)Tj
1.9162 0 Td
(with)Tj
2.0012 0 Td
(high)Tj
2.0069 0 Td
(and)Tj
1.7292 0 Td
(low)Tj
1.6554 0 Td
(expression)Tj
4.4674 0 Td
(of)Tj
-34.333 -1.3039 Td
(ANO1.)Tj
3.0841 0 Td
(To)Tj
1.3436 0 Td
(separate)Tj
3.4809 0 Td
(high)Tj
2.0069 0 Td
(and)Tj
1.7291 0 Td
(low)Tj
1.6611 0 Td
(levels)Tj
2.3697 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
2.863 0 Td
(expressing)Tj
4.4277 0 Td
(PC-3)Tj
2.2677 0 Td
(cells,)Tj
2.143 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(stably)Tj
2.5228 0 Td
(trans-)Tj
-34.9566 -1.2982 Td
(fected)Tj
2.6022 0 Td
(with)Tj
2.0013 0 Td
(the)Tj
1.4683 0 Td
(halide)Tj
2.6418 0 Td
(sensor)Tj
2.7949 0 Td
(YFP-H148Q/I152L/F46)Tj
9.4563 0 Td
(L)Tj
0.7541 0 Td
(and)Tj
1.7348 0 Td
(then)Tj
2.0182 0 Td
(the)Tj
1.4683 0 Td
(cells)Tj
1.9163 0 Td
(were)Tj
2.1259 0 Td
(plated)Tj
2.6646 0 Td
(on)Tj
1.2756 0 Td
(96)Tj
-34.9226 -1.2983 Td
(well)Tj
1.8199 0 Td
(plates)Tj
2.5001 0 Td
(at)Tj
0.9524 0 Td
(1)Tj
0.6974 0 Td
(cell)Tj
1.5589 0 Td
(per)Tj
1.5307 0 Td
(well.)Tj
2.0466 0 Td
(After)Tj
2.2847 0 Td
(two)Tj
1.7065 0 Td
(weeks)Tj
2.6078 0 Td
(of)Tj
1.0148 0 Td
(incubation,)Tj
4.7792 0 Td
(high)Tj
2.0069 0 Td
(and)Tj
1.7291 0 Td
(low)Tj
1.6555 0 Td
(levels)Tj
2.3754 0 Td
(of)Tj
1.0148 0 Td
(ANO1)Tj
-32.2807 -1.2983 Td
(expressing)Tj
4.4221 0 Td
(PC-3)Tj
2.2677 0 Td
(cells)Tj
1.9217 0 Td
(were)Tj
2.1203 0 Td
(separated)Tj
4.0139 0 Td
(by)Tj
1.1792 0 Td
(YFP)Tj
1.9389 0 Td
(quenching)Tj
4.4617 0 Td
(assay.)Tj
2.4888 0 Td
(ANO1)Tj
2.8629 0 Td
(was)Tj
1.6952 0 Td
(activated)Tj
3.7813 0 Td
(by)Tj
1.1849 0 Td
(100)Tj
/F11 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
(M)Tj
-35.9827 -1.3039 Td
(ATP.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 613.8141 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cell)Tj
1.9134 0 Td
(based)Tj
2.9102 0 Td
(screening)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 596.8062 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(ANO1)Tj
2.8573 0 Td
(and)Tj
1.7291 0 Td
(YFP)Tj
1.9332 0 Td
(expressing)Tj
4.4163 0 Td
(FRT)Tj
1.9786 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
2.1203 0 Td
(plated)Tj
2.6646 0 Td
(in)Tj
1.0318 0 Td
(96-well)Tj
3.1181 0 Td
(black-walled)Tj
5.2327 0 Td
(microplates)Tj
4.8869 0 Td
(\(Corning)Tj
-33.8851 -1.2982 Td
(Inc.,)Tj
1.9842 0 Td
(Corning,)Tj
3.8154 0 Td
(NY\))Tj
1.9275 0 Td
(at)Tj
0.9525 0 Td
(a)Tj
0.6519 0 Td
(density)Tj
3.1011 0 Td
(of)Tj
1.0148 0 Td
(20,000)Tj
2.8233 0 Td
(cells)Tj
1.9219 0 Td
(per)Tj
1.5307 0 Td
(well.)Tj
2.0466 0 Td
(Assays)Tj
2.8743 0 Td
(were)Tj
2.126 0 Td
(done)Tj
2.2223 0 Td
(using)Tj
2.3924 0 Td
(FLUOstar)Tj
-31.3849 -1.3039 Td
(Omega)Tj
3.1067 0 Td
(microplate)Tj
4.5297 0 Td
(reader)Tj
2.7723 0 Td
(\(BMG)Tj
2.7439 0 Td
(Labtech,)Tj
3.6 0 Td
(Ortenberg,)Tj
4.6034 0 Td
(Germany\))Tj
4.3257 0 Td
(and)Tj
1.7291 0 Td
(MARS)Tj
2.897 0 Td
(Data)Tj
2.1203 0 Td
(Analysis)Tj
-32.4281 -1.2983 Td
(Software)Tj
3.702 0 Td
(\(BMG)Tj
2.7439 0 Td
(Labtech\))Tj
3.7134 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(in)Tj
1.0374 0 Td
(our)Tj
1.6271 0 Td
(previous)Tj
3.6454 0 Td
(study)Tj
2.3924 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(].)Tj
2.0749 0 Td
(Briefly,)Tj
3.0784 0 Td
(each)Tj
2.0296 0 Td
(well)Tj
1.8199 0 Td
(of)Tj
1.0148 0 Td
(96-well)Tj
-33.9475 -1.2983 Td
(plate)Tj
2.1373 0 Td
(was)Tj
1.7007 0 Td
(washed)Tj
3.1748 0 Td
(3)Tj
0.6973 0 Td
(times)Tj
2.3925 0 Td
(in)Tj
1.0318 0 Td
(PBS)Tj
1.8368 0 Td
(\(200)Tj
/F11 1 Tf
1.9842 0 Td
[()]TJ
/F7 1 Tf
(L/wash\))Tj
3.9458 0 Td
(and)Tj
1.7292 0 Td
(test)Tj
1.5987 0 Td
(compounds)Tj
4.9606 0 Td
(were)Tj
2.1203 0 Td
(added)Tj
2.6646 0 Td
(to)Tj
1.0261 0 Td
(each)Tj
2.0239 0 Td
(well)Tj
1.8198 0 Td
(at)Tj
-36.8444 -1.3039 Td
(25)Tj
/F11 1 Tf
1.1678 0 Td
[()]TJ
/F7 1 Tf
(M)Tj
1.6214 0 Td
(final)Tj
2.0013 0 Td
(concentration.)Tj
6.0491 0 Td
(After)Tj
2.2847 0 Td
(10)Tj
1.1678 0 Td
(minutes)Tj
3.4696 0 Td
(incubation)Tj
4.5468 0 Td
(at)Tj
0.9524 0 Td
(37C,)Tj
2.4548 0 Td
(96-well)Tj
3.1181 0 Td
(plates)Tj
2.5001 0 Td
(were)Tj
2.126 0 Td
(trans-)Tj
-33.4599 -1.2982 Td
(ferred)Tj
2.6248 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.4683 0 Td
(plate)Tj
2.1373 0 Td
(reader)Tj
2.7723 0 Td
(for)Tj
1.3889 0 Td
(fluorescence)Tj
5.1874 0 Td
(assay.)Tj
2.4888 0 Td
(Each)Tj
2.1713 0 Td
(well)Tj
1.8199 0 Td
(was)Tj
1.7007 0 Td
(assayed)Tj
3.2145 0 Td
(individually)Tj
4.9889 0 Td
(for)Tj
1.389 0 Td
(ANO1-)Tj
-34.3783 -1.2983 Td
(mediated)Tj
3.9401 0 Td
(I)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 242.8157 510.2362 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 247.6346 505.8141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(influx)Tj
2.5625 0 Td
(by)Tj
1.1792 0 Td
(monitoring)Tj
4.8245 0 Td
(YFP)Tj
1.9389 0 Td
(fluorescence)Tj
5.1873 0 Td
(continuously)Tj
5.4369 0 Td
(\(400)Tj
1.9842 0 Td
(ms)Tj
1.4003 0 Td
(per)Tj
1.5307 0 Td
(point\))Tj
2.6986 0 Td
(for)Tj
1.3889 0 Td
(2)Tj
0.6974 0 Td
(s)Tj
-35.5915 -1.3039 Td
(\(baseline\),)Tj
4.337 0 Td
(then)Tj
2.0181 0 Td
(140)Tj
1.6441 0 Td
(mM)Tj
1.9332 0 Td
(I)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 302.7401 497.2535 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 307.559 492.7748 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(solution)Tj
3.4809 0 Td
(containing)Tj
4.5297 0 Td
(200)Tj
/F11 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
(M)Tj
1.6214 0 Td
(ATP)Tj
2.0749 0 Td
(was)Tj
1.7008 0 Td
(added)Tj
2.6589 0 Td
(at)Tj
0.9524 0 Td
(2)Tj
0.6974 0 Td
(s)Tj
0.5782 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(YFP)Tj
-34.4406 -1.2982 Td
(fluorescence)Tj
5.1873 0 Td
(was)Tj
1.6951 0 Td
(recorded)Tj
3.7984 0 Td
(for)Tj
1.3832 0 Td
(6)Tj
0.6974 0 Td
(s.)Tj
0.805 0 Td
(Initial)Tj
2.6249 0 Td
(iodide)Tj
2.7382 0 Td
(influx)Tj
2.5625 0 Td
(rate)Tj
1.7462 0 Td
(was)Tj
1.7007 0 Td
(determined)Tj
4.8529 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(initial)Tj
-33.4655 -1.2983 Td
(slope)Tj
2.2733 0 Td
(of)Tj
1.0148 0 Td
(fluorescence)Tj
5.1874 0 Td
(decrease,)Tj
3.8323 0 Td
(by)Tj
1.1849 0 Td
(nonlinear)Tj
4.1102 0 Td
(regression,)Tj
4.5468 0 Td
(following)Tj
3.9798 0 Td
(infusion)Tj
3.5319 0 Td
(of)Tj
1.0148 0 Td
(iodide)Tj
2.7439 0 Td
(with)Tj
1.9956 0 Td
(ATP.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 438.1795 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Short-circuit)Tj
5.4945 0 Td
(current)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 421.2283 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Snapwell)Tj
3.7927 0 Td
(inserts)Tj
2.8516 0 Td
(containing)Tj
4.5298 0 Td
(ANO1-expressing)Tj
7.4153 0 Td
(FRT)Tj
1.9786 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.126 0 Td
(mounted)Tj
3.8721 0 Td
(in)Tj
1.0318 0 Td
(Ussing)Tj
2.9593 0 Td
(chambers)Tj
-32.4791 -1.3039 Td
(\(Physiologic)Tj
5.142 0 Td
(Instruments,)Tj
5.3461 0 Td
(San)Tj
1.6781 0 Td
(Diego,)Tj
2.8459 0 Td
(CA\).)Tj
2.1317 0 Td
(The)Tj
1.7801 0 Td
(basolateral)Tj
4.4844 0 Td
(bath)Tj
1.9842 0 Td
(was)Tj
1.7008 0 Td
(filled)Tj
2.2167 0 Td
(with)Tj
2.0012 0 Td
(HCO3)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 539.6598 412.6677 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 542.2677 408.1889 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-buff-)Tj
-34.2252 -1.2982 Td
(ered)Tj
1.9672 0 Td
(solution)Tj
3.4809 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(apical)Tj
2.5455 0 Td
(bath)Tj
1.9842 0 Td
(was)Tj
1.7008 0 Td
(filled)Tj
2.2167 0 Td
(with)Tj
2.0012 0 Td
(a)Tj
0.652 0 Td
(half-Cl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 425.0267 399.6283 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 429.8456 395.2062 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(solution.)Tj
3.7076 0 Td
(The)Tj
1.7802 0 Td
(basolateral)Tj
4.4844 0 Td
(membrane)Tj
-32.9553 -1.2982 Td
(was)Tj
1.6951 0 Td
(permeabilized)Tj
5.862 0 Td
(with)Tj
2.0013 0 Td
(250)Tj
/F11 1 Tf
1.644 0 Td
[()]TJ
/F7 1 Tf
(g/mL)Tj
2.88 0 Td
(amphotericin)Tj
5.6239 0 Td
(B.)Tj
1.0262 0 Td
(The)Tj
1.7801 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.126 0 Td
(bathed)Tj
2.9366 0 Td
(for)Tj
1.3833 0 Td
(a)Tj
0.6577 0 Td
(20)Tj
1.1678 0 Td
(min)Tj
1.8595 0 Td
(stabili-)Tj
-34.5653 -1.3039 Td
(zation)Tj
2.6872 0 Td
(period)Tj
2.8346 0 Td
(and)Tj
1.7292 0 Td
(aerated)Tj
3.1294 0 Td
(with)Tj
2.0012 0 Td
(95%)Tj
1.9276 0 Td
(O)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 350.5889 367.5401 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 356.3149 369.1842 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(/)Tj
0.5499 0 Td
(5%CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 388.233 367.5401 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 394.0157 369.1842 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(at)Tj
0.9524 0 Td
(37C.)Tj
2.4491 0 Td
(Luteolin)Tj
3.5773 0 Td
(and)Tj
1.7291 0 Td
(kaempferol)Tj
4.7509 0 Td
(were)Tj
-32.8589 -1.2982 Td
(applied)Tj
3.1521 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(apical)Tj
2.5399 0 Td
(and)Tj
1.7348 0 Td
(basolateral)Tj
4.4787 0 Td
(bath)Tj
1.9899 0 Td
(solution)Tj
3.4808 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(ATP)Tj
2.0749 0 Td
(was)Tj
1.7008 0 Td
(added)Tj
2.6589 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(apical)Tj
2.5454 0 Td
(bath)Tj
-35.0925 -1.2983 Td
(solution)Tj
3.4809 0 Td
(to)Tj
1.0261 0 Td
(induce)Tj
2.9367 0 Td
(intracellular)Tj
5.057 0 Td
(calcium)Tj
3.3618 0 Td
(increase.)Tj
3.702 0 Td
(Apical)Tj
2.7892 0 Td
(membrane)Tj
4.5695 0 Td
(currents)Tj
3.5432 0 Td
(were)Tj
2.1203 0 Td
(measured)Tj
-32.5867 -1.3039 Td
(with)Tj
1.9955 0 Td
(an)Tj
1.2019 0 Td
(EVC4000)Tj
4.0535 0 Td
(Multi-Channel)Tj
6.1909 0 Td
(V/I)Tj
1.5874 0 Td
(Clamp)Tj
2.8969 0 Td
(\(World)Tj
3.1804 0 Td
(Precision)Tj
3.9458 0 Td
(Instruments,)Tj
5.3461 0 Td
(Sarasota,)Tj
3.7587 0 Td
(FL\))Tj
1.6271 0 Td
(and)Tj
-35.7842 -1.2983 Td
(recorded)Tj
3.7927 0 Td
(using)Tj
2.3924 0 Td
(PowerLab)Tj
4.2406 0 Td
(4/35)Tj
1.9786 0 Td
(\(AD)Tj
1.9786 0 Td
(Instruments,)Tj
5.346 0 Td
(Castle)Tj
2.6475 0 Td
(Hill,)Tj
1.9729 0 Td
(Australia\).)Tj
4.405 0 Td
(Data)Tj
2.126 0 Td
(were)Tj
2.1203 0 Td
(collected)Tj
-33.0006 -1.2982 Td
(and)Tj
1.7291 0 Td
(analyzed)Tj
3.702 0 Td
(with)Tj
1.9956 0 Td
(ADInstruments)Tj
6.5423 0 Td
(acquisition)Tj
4.6261 0 Td
(software)Tj
3.5886 0 Td
(Labchart)Tj
3.753 0 Td
(Pro)Tj
1.6554 0 Td
(7)Tj
0.6973 0 Td
(software.)Tj
3.8098 0 Td
(The)Tj
1.7858 0 Td
(sampling)Tj
-33.885 -1.3039 Td
(rate)Tj
1.7461 0 Td
(was)Tj
1.7008 0 Td
(4)Tj
0.6916 0 Td
(Hz.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 262.6015 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Intracellular)Tj
5.315 0 Td
(calcium)Tj
3.5716 0 Td
(measurement)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 245.5937 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Intracellular)Tj
5.1306 0 Td
(calcium)Tj
3.3619 0 Td
(levels)Tj
2.3754 0 Td
(were)Tj
2.1203 0 Td
(monitored)Tj
4.4901 0 Td
(using)Tj
2.3924 0 Td
(Fluo-4)Tj
2.8516 0 Td
(NW)Tj
1.9219 0 Td
(calcium)Tj
3.3619 0 Td
(assay)Tj
2.2677 0 Td
(kit)Tj
1.2812 0 Td
(\(Invitrogen,)Tj
-31.555 -1.2982 Td
(Carlsbad,)Tj
3.9911 0 Td
(CA\))Tj
1.9049 0 Td
(as)Tj
1.0148 0 Td
(per)Tj
1.5307 0 Td
(the)Tj
1.4683 0 Td
(manufacturer's)Tj
6.2645 0 Td
(protocol)Tj
3.6 0 Td
(in)Tj
1.0375 0 Td
(FRT)Tj
1.9786 0 Td
(and)Tj
1.7291 0 Td
(PC-3)Tj
2.2677 0 Td
(cells.)Tj
2.1486 0 Td
(Briefly,)Tj
3.0728 0 Td
(the)Tj
1.474 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
-35.3988 -1.2983 Td
(cultured)Tj
3.5659 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.6519 0 Td
(96)Tj
1.1679 0 Td
(well)Tj
1.8198 0 Td
(plate)Tj
2.143 0 Td
(and)Tj
1.7291 0 Td
(incubated)Tj
4.1783 0 Td
(with)Tj
1.9956 0 Td
(100)Tj
/F11 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2643 0 Td
(assay)Tj
2.262 0 Td
(buffer)Tj
2.6249 0 Td
(including)Tj
4.0421 0 Td
(Fluo-4)Tj
2.8573 0 Td
(NW.)Tj
2.1487 0 Td
(After)Tj
-35.138 -1.3039 Td
(1)Tj
0.6916 0 Td
(hour)Tj
2.1543 0 Td
(incubation,)Tj
4.7735 0 Td
(Fluo-4)Tj
2.8573 0 Td
(fluorescence)Tj
5.1874 0 Td
(was)Tj
1.7008 0 Td
(measured)Tj
4.1045 0 Td
(with)Tj
2.0013 0 Td
(a)Tj
0.6519 0 Td
(FLUOstar)Tj
4.235 0 Td
(Omega)Tj
3.1067 0 Td
(microplate)Tj
-31.4643 -1.2983 Td
(reader)Tj
2.7722 0 Td
(equipped)Tj
3.9288 0 Td
(with)Tj
1.9956 0 Td
(syringe)Tj
3.1124 0 Td
(pumps)Tj
2.965 0 Td
(and)Tj
1.7292 0 Td
(custom)Tj
3.1577 0 Td
(Fluo-4)Tj
2.8517 0 Td
(excitation/emission)Tj
8.039 0 Td
(filters)Tj
2.4774 0 Td
(\(485/538)Tj
-33.029 -1.2982 Td
(nm\).)Tj
2.1543 0 Td
(Intracellular)Tj
5.1306 0 Td
(calcium)Tj
3.3619 0 Td
(increase)Tj
3.4696 0 Td
(was)Tj
1.7008 0 Td
(induced)Tj
3.4639 0 Td
(by)Tj
1.1849 0 Td
(application)Tj
4.6771 0 Td
(of)Tj
1.0148 0 Td
(100)Tj
/F11 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(M)Tj
1.6158 0 Td
(ATP.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 151.9937 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Patch-clamp)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 134.9858 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Patch-clamp)Tj
5.2497 0 Td
(recordings)Tj
4.49 0 Td
(were)Tj
2.1203 0 Td
(performed)Tj
4.4731 0 Td
(on)Tj
1.2756 0 Td
(ANO1-expressing)Tj
7.4153 0 Td
(FRT)Tj
1.9786 0 Td
(cells)Tj
1.9219 0 Td
(using)Tj
2.3867 0 Td
(the)Tj
1.4741 0 Td
(whole-cell)Tj
-32.7853 -1.2982 Td
(recording)Tj
4.1215 0 Td
(configuration.)Tj
5.9017 0 Td
(The)Tj
1.7801 0 Td
(bath)Tj
1.9899 0 Td
(solution)Tj
3.481 0 Td
(contained)Tj
4.2009 0 Td
(\(in)Tj
1.372 0 Td
(mM\):)Tj
2.5001 0 Td
(140)Tj
1.6441 0 Td
(NMDG-Cl,)Tj
4.7905 0 Td
(1)Tj
0.6916 0 Td
(CaCl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 544.8188 120.359 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 548.3905 122.0031 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4478 0 Td
(1)Tj
-35.2853 -1.2982 Td
(MgCl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 222.6897 107.3196 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 226.2047 109.0204 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4478 0 Td
(10)Tj
1.1736 0 Td
(glucose)Tj
3.1691 0 Td
(and)Tj
1.7348 0 Td
(10)Tj
1.1678 0 Td
(HEPES)Tj
3.1635 0 Td
(\(pH)Tj
1.8425 0 Td
(7.4\).)Tj
1.9615 0 Td
(The)Tj
1.7859 0 Td
(pipette)Tj
2.965 0 Td
(solution)Tj
3.4809 0 Td
(contained)Tj
4.2009 0 Td
(\(in)Tj
1.372 0 Td
(mM\):)Tj
2.5001 0 Td
(130)Tj
1.6441 0 Td
(CsCl,)Tj
-35.2286 -1.3039 Td
(0.5)Tj
1.3946 0 Td
(EGTA,)Tj
3.0273 0 Td
(1)Tj
0.6973 0 Td
(MgCl)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 273.8267 94.337 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 277.3984 95.9811 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4478 0 Td
(1)Tj
0.6917 0 Td
(Tris-ATP,)Tj
4.2633 0 Td
(and)Tj
1.7291 0 Td
(10)Tj
1.1735 0 Td
(HEPES)Tj
3.1578 0 Td
(\(pH)Tj
1.8482 0 Td
(7.2\).)Tj
1.9615 0 Td
(Pipettes)Tj
3.3676 0 Td
(were)Tj
2.1203 0 Td
(pulled)Tj
2.7099 0 Td
(from)Tj
2.2053 0 Td
(borosili-)Tj
-33.4145 -1.2982 Td
(cate)Tj
1.7972 0 Td
(glass)Tj
2.0863 0 Td
(and)Tj
1.7347 0 Td
(had)Tj
1.7065 0 Td
(resistances)Tj
4.4956 0 Td
(of)Tj
1.0148 0 Td
(35)Tj
1.6838 0 Td
(M)Tj
/F12 1 Tf
(O)Tj
/F7 1 Tf
1.8311 0 Td
(after)Tj
2.0353 0 Td
(fire)Tj
1.5704 0 Td
(polishing.)Tj
4.1499 0 Td
(Seal)Tj
1.8028 0 Td
(resistances)Tj
4.4901 0 Td
(were)Tj
2.1203 0 Td
(between)Tj
3.5319 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 453.2598 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Downre)Tj
3.3803 0 Td
(gulation)Tj
3.6496 0 Td
(of)Tj
1.0418 0 Td
(ANO1)Tj
2.9126 0 Td
(by)Tj
1.2543 0 Td
(luteolin)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01749)Tj
4.089 0 Td
(35)Tj
0 g
2.0906 0 Td
(March)Tj
2.9622 0 Td
(31,)Tj
1.5874 0 Td
(2017)Tj
34.3562 0 Td
(3)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
46 0 obj
<>stream
endstream
endobj
47 0 obj
<>stream
endstream
endobj
48 0 obj
<>stream
endstream
endobj
49 0 obj
<>stream
endstream
endobj
50 0 obj
<>stream
BT
66.402 0 Td
(13)Tj
ET
Q
endstream
endobj
51 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 10 0 R/Annots 52 0 R/Contents 53 0 R/TrimBox[0 0 612 792]>>
endobj
52 0 obj
[54 0 R 55 0 R]
endobj
54 0 obj
<>/Border[0 0 0]/Dest(Rpone.0174935.ref018)>>
endobj
55 0 obj
<>/Border[0 0 0]/A 56 0 R>>
endobj
56 0 obj
<>
endobj
53 0 obj
[57 0 R 58 0 R 59 0 R 60 0 R 61 0 R 62 0 R]
endobj
57 0 obj
<>stream
q
0.83 0.64 0.02 0 k
428.485 309.5433 m
437.9528 309.5433 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(and)Tj
1.7291 0 Td
(10)Tj
1.1735 0 Td
(G)Tj
/F12 1 Tf
(O)Tj
/F7 1 Tf
(.)Tj
1.8765 0 Td
(The)Tj
1.7802 0 Td
(liquid)Tj
2.5625 0 Td
(junction)Tj
3.6 0 Td
(potential)Tj
3.7587 0 Td
(\(~2.4)Tj
2.313 0 Td
(mV\))Tj
2.075 0 Td
(was)Tj
1.7008 0 Td
(not)Tj
1.576 0 Td
(corrected.)Tj
4.2123 0 Td
(After)Tj
2.2847 0 Td
(establishing)Tj
4.9266 0 Td
(the)Tj
-35.5689 -1.3039 Td
(whole-cell)Tj
4.2972 0 Td
(configuration,)Tj
5.8961 0 Td
(whole-cell)Tj
4.3029 0 Td
(capacitance)Tj
4.8473 0 Td
(and)Tj
1.7291 0 Td
(series)Tj
2.4264 0 Td
(resistance)Tj
4.1329 0 Td
(were)Tj
2.1203 0 Td
(compensated)Tj
5.5049 0 Td
(with)Tj
-35.2571 -1.2982 Td
(the)Tj
1.4683 0 Td
(amplifier)Tj
3.8551 0 Td
(circuitry.)Tj
3.8494 0 Td
(After)Tj
2.2847 0 Td
(establishing)Tj
4.9266 0 Td
(the)Tj
1.4683 0 Td
(whole-cell)Tj
4.2973 0 Td
(configuration,)Tj
5.9017 0 Td
(ANO1)Tj
2.863 0 Td
(was)Tj
1.7008 0 Td
(activated)Tj
3.7814 0 Td
(by)Tj
-36.3966 -1.2983 Td
(ATP)Tj
2.0749 0 Td
(\(100)Tj
/F11 1 Tf
1.9842 0 Td
[()]TJ
/F7 1 Tf
(M\).)Tj
2.1884 0 Td
(Whole-cell)Tj
4.5751 0 Td
(currents)Tj
3.5489 0 Td
(were)Tj
2.1203 0 Td
(elicited)Tj
3.0954 0 Td
(by)Tj
1.1849 0 Td
(applying)Tj
3.668 0 Td
(hyperpolarizing)Tj
6.5537 0 Td
(and)Tj
1.7291 0 Td
(depolariz-)Tj
-32.7229 -1.2983 Td
(ing)Tj
1.5023 0 Td
(voltage)Tj
3.0444 0 Td
(pulses)Tj
2.6589 0 Td
(from)Tj
2.2053 0 Td
(a)Tj
0.652 0 Td
(holding)Tj
3.3108 0 Td
(potential)Tj
3.7587 0 Td
(of)Tj
1.0148 0 Td
(0)Tj
0.6974 0 Td
(mV)Tj
1.7347 0 Td
(to)Tj
1.0262 0 Td
(potentials)Tj
4.1215 0 Td
(between)Tj
3.5263 0 Td
(-80)Tj
1.525 0 Td
(mV)Tj
1.7348 0 Td
(and)Tj
1.7292 0 Td
(+80)Tj
1.7518 0 Td
(mV)Tj
-35.9941 -1.3039 Td
(in)Tj
1.0318 0 Td
(steps)Tj
2.1826 0 Td
(of)Tj
1.0148 0 Td
(20)Tj
1.1736 0 Td
(mV.)Tj
1.9558 0 Td
(Recordings)Tj
4.7395 0 Td
(were)Tj
2.1203 0 Td
(made)Tj
2.4265 0 Td
(at)Tj
0.9524 0 Td
(room)Tj
2.4265 0 Td
(temperature)Tj
5.125 0 Td
(using)Tj
2.3924 0 Td
(an)Tj
1.2019 0 Td
(Axopatch-200B)Tj
-28.7431 -1.2982 Td
(\(Axon)Tj
2.7609 0 Td
(Instruments,)Tj
5.3461 0 Td
(Union)Tj
2.8289 0 Td
(City,)Tj
2.126 0 Td
(CA\).)Tj
2.1373 0 Td
(Currents)Tj
3.7814 0 Td
(were)Tj
2.126 0 Td
(digitized)Tj
3.6737 0 Td
(and)Tj
1.7291 0 Td
(analyzed)Tj
3.702 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.652 0 Td
(Digidata)Tj
-33.2558 -1.2983 Td
(1440A)Tj
2.8006 0 Td
(converter)Tj
4.0365 0 Td
(\(Axon)Tj
2.7609 0 Td
(Instruments\),)Tj
5.6806 0 Td
(and)Tj
1.7291 0 Td
(pCLAMP)Tj
4.0705 0 Td
(10.2)Tj
1.8652 0 Td
(software)Tj
3.5887 0 Td
(\(Molecular)Tj
4.6261 0 Td
(Devices,)Tj
3.5319 0 Td
(Sunny-)Tj
-34.6901 -1.3039 Td
(vale,)Tj
2.0012 0 Td
(CA\).)Tj
2.1316 0 Td
(Currents)Tj
3.7871 0 Td
(were)Tj
2.126 0 Td
(low-pass)Tj
3.6793 0 Td
(filtered)Tj
3.0671 0 Td
(at)Tj
0.9524 0 Td
(1)Tj
0.6917 0 Td
(kHz)Tj
1.8992 0 Td
(and)Tj
1.7291 0 Td
(sampled)Tj
3.5489 0 Td
(at)Tj
0.9525 0 Td
(5)Tj
0.6973 0 Td
(kHz.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 574.8094 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cell)Tj
1.9134 0 Td
(proliferation)Tj
5.3669 0 Td
(assays)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 557.8015 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(PC-3)Tj
2.262 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
2.1203 0 Td
(seeded)Tj
2.9027 0 Td
(to)Tj
1.0204 0 Td
(a)Tj
0.6463 0 Td
(confluency)Tj
4.6091 0 Td
(of)Tj
1.0091 0 Td
(~30%)Tj
2.5059 0 Td
(on)Tj
1.2699 0 Td
(Day)Tj
1.8425 0 Td
(1)Tj
0.6916 0 Td
(in)Tj
1.0318 0 Td
(96-well)Tj
3.1181 0 Td
(plates.)Tj
2.7213 0 Td
(The)Tj
1.7744 0 Td
(cells)Tj
1.9163 0 Td
(were)Tj
2.1203 0 Td
(incu-)Tj
-35.4782 -1.2982 Td
(bated)Tj
2.4094 0 Td
(for)Tj
1.3833 0 Td
(24)Tj
1.1735 0 Td
(hours)Tj
2.5172 0 Td
(to)Tj
1.0204 0 Td
(reach)Tj
2.4038 0 Td
(~50%)Tj
2.5058 0 Td
(confluence)Tj
4.5808 0 Td
(and)Tj
1.7291 0 Td
(treated)Tj
2.999 0 Td
(with)Tj
1.9956 0 Td
(different)Tj
3.6567 0 Td
(concentrations)Tj
6.1795 0 Td
(\(3,)Tj
1.2642 0 Td
(10,)Tj
-35.8183 -1.3039 Td
(30,)Tj
1.3946 0 Td
(100)Tj
/F11 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(M\))Tj
1.9559 0 Td
(of)Tj
1.0148 0 Td
(luteolin)Tj
3.2882 0 Td
(and)Tj
1.7291 0 Td
(kaempferol,)Tj
4.972 0 Td
(and)Tj
1.7291 0 Td
(an)Tj
1.2019 0 Td
(equal)Tj
2.3584 0 Td
(amount)Tj
3.3562 0 Td
(of)Tj
1.0148 0 Td
(DMSO)Tj
3.0727 0 Td
(was)Tj
1.6951 0 Td
(added)Tj
2.6646 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(all)Tj
-35.5915 -1.2983 Td
(control.)Tj
3.3448 0 Td
(After)Tj
2.2847 0 Td
(24)Tj
1.1736 0 Td
(hour)Tj
2.1543 0 Td
(elapsed,)Tj
3.3789 0 Td
(the)Tj
1.4683 0 Td
(influence)Tj
3.9174 0 Td
(of)Tj
1.0148 0 Td
(luteolin)Tj
3.2825 0 Td
(and)Tj
1.7292 0 Td
(kaempferol)Tj
4.7508 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4683 0 Td
(cell)Tj
1.5591 0 Td
(viability)Tj
-32.8023 -1.2983 Td
(was)Tj
1.6951 0 Td
(estimated)Tj
4.0705 0 Td
(by)Tj
1.1792 0 Td
(MTS)Tj
2.1997 0 Td
(assay.)Tj
2.4831 0 Td
(Briefly,)Tj
3.0727 0 Td
(the)Tj
1.4684 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1147 0 Td
(reincubated)Tj
4.9719 0 Td
(with)Tj
1.9956 0 Td
(MTS)Tj
2.194 0 Td
(for)Tj
1.3833 0 Td
(1)Tj
0.6973 0 Td
(hour.)Tj
2.3754 0 Td
(The)Tj
1.7802 0 Td
(solu-)Tj
-35.6029 -1.3039 Td
(ble)Tj
1.3946 0 Td
(formazan)Tj
4.0365 0 Td
(produced)Tj
4.0478 0 Td
(by)Tj
1.1793 0 Td
(cellular)Tj
3.1407 0 Td
(reduction)Tj
4.1102 0 Td
(of)Tj
1.0148 0 Td
(MTS)Tj
2.1997 0 Td
(was)Tj
1.7008 0 Td
(quantified)Tj
4.3086 0 Td
(by)Tj
1.1849 0 Td
(measuring)Tj
4.4333 0 Td
(the)Tj
1.474 0 Td
(absor-)Tj
-34.2252 -1.2982 Td
(bance)Tj
2.5568 0 Td
(at)Tj
0.9524 0 Td
(490)Tj
1.6441 0 Td
(nm)Tj
1.5874 0 Td
(with)Tj
2.0012 0 Td
(Infinite)Tj
3.1975 0 Td
(M200)Tj
2.5398 0 Td
(\(Tecan,)Tj
3.2258 0 Td
(Gro)Tj
0 Tc
1.1509 0 Td
()Tj
-0.004 Tc
0.4422 0 Td
(dig,)Tj
1.7064 0 Td
(Austria\))Tj
3.5036 0 Td
(microplate)Tj
4.5298 0 Td
(reader.)Tj
2.999 0 Td
(MTS)Tj
2.194 0 Td
(assay)Tj
-34.2309 -1.2983 Td
(was)Tj
1.6951 0 Td
(done)Tj
2.228 0 Td
(using)Tj
2.3924 0 Td
(CellTiter)Tj
3.7701 0 Td
(96)Tj
1.1678 0 Td
(Aqueous)Tj
3.7587 0 Td
(One)Tj
1.9389 0 Td
(Solution)Tj
3.6 0 Td
(Cell)Tj
1.7972 0 Td
(Proliferation)Tj
5.312 0 Td
(Assay)Tj
2.5115 0 Td
(kit)Tj
1.2813 0 Td
(\(Promega,)Tj
-31.453 -1.2983 Td
(Madison,)Tj
3.9798 0 Td
(WI\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 425.1968 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Scratch)Tj
3.5764 0 Td
(wound)Tj
3.1275 0 Td
(healing)Tj
3.3922 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 408.1889 cm
BT
/F7 1 Tf
1 TL
-0.01 Tc
0 0 Td
(PC-3)Tj
2.245 0 Td
(cells)Tj
1.8878 0 Td
(were)Tj
2.0977 0 Td
(cultured)Tj
3.5206 0 Td
(in)Tj
1.0204 0 Td
(a)Tj
0.652 0 Td
(96-well)Tj
3.0784 0 Td
(plate)Tj
2.109 0 Td
(until)Tj
2.0806 0 Td
(they)Tj
1.8992 0 Td
(reached)Tj
3.3108 0 Td
(~80%)Tj
2.4832 0 Td
(confluence)Tj
4.5184 0 Td
(as)Tj
1.0034 0 Td
(a)Tj
0.6463 0 Td
(monolayer)Tj
-32.5528 -1.2982 Td
(and)Tj
1.7121 0 Td
(then)Tj
1.9955 0 Td
(the)Tj
1.4514 0 Td
(cell)Tj
1.5363 0 Td
(layer)Tj
2.1147 0 Td
(was)Tj
1.6837 0 Td
(wounded)Tj
3.9232 0 Td
(using)Tj
2.3584 0 Td
(a)Tj
0.6519 0 Td
(96-Well)Tj
3.3619 0 Td
(WoundMaker)Tj
5.8507 0 Td
(\(Essen)Tj
2.7893 0 Td
(BioScience,)Tj
4.7168 0 Td
(MI\))Tj
1.7745 0 Td
(and)Tj
-35.9204 -1.2983 Td
(washed)Tj
3.1407 0 Td
(twice)Tj
2.2847 0 Td
(with)Tj
1.9729 0 Td
(fresh)Tj
2.1657 0 Td
(serum)Tj
2.6929 0 Td
(free)Tj
1.7008 0 Td
(media.)Tj
2.8799 0 Td
(The)Tj
1.7688 0 Td
(cells)Tj
1.8879 0 Td
(were)Tj
2.0976 0 Td
(incubated)Tj
4.1273 0 Td
(with)Tj
1.9729 0 Td
(serum)Tj
2.6928 0 Td
(free)Tj
1.7008 0 Td
(medium,)Tj
-33.0857 -1.3039 Td
(and)Tj
1.7121 0 Td
(images)Tj
2.9536 0 Td
(of)Tj
1.0035 0 Td
(the)Tj
1.4513 0 Td
(wounds)Tj
3.3392 0 Td
(were)Tj
2.0977 0 Td
(taken)Tj
2.3924 0 Td
(using)Tj
2.3584 0 Td
(the)Tj
1.4513 0 Td
(IncuCyte)Tj
3.8551 0 Td
(ZOOM)Tj
3.1805 0 Td
(\(Essen)Tj
2.7892 0 Td
(BioScience,)Tj
4.7169 0 Td
(MI\).)Tj
1.9955 0 Td
(The)Tj
-35.2967 -1.2983 Td
(wound)Tj
2.982 0 Td
(recovery)Tj
3.6056 0 Td
(properties)Tj
4.218 0 Td
(of)Tj
1.0034 0 Td
(test)Tj
1.5761 0 Td
(compounds)Tj
4.9039 0 Td
(were)Tj
2.0976 0 Td
(analyzed)Tj
3.6567 0 Td
(by)Tj
1.1678 0 Td
(the)Tj
1.4514 0 Td
(IncuCyte)Tj
3.8551 0 Td
(software.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 327.6283 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Immunoblot)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 310.6204 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Immunoblotting)Tj
6.9164 0 Td
(was)Tj
1.7008 0 Td
(performed)Tj
4.4674 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(previously)Tj
4.354 0 Td
([)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(].)Tj
2.0749 0 Td
(FRT)Tj
1.9786 0 Td
(and)Tj
1.7348 0 Td
(PC-3)Tj
2.2677 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
2.126 0 Td
(lysed)Tj
-34.6051 -1.3039 Td
(with)Tj
1.9955 0 Td
(cell)Tj
1.5591 0 Td
(lysis)Tj
1.9105 0 Td
(buffer)Tj
2.6249 0 Td
(\(50)Tj
1.5137 0 Td
(mM)Tj
1.9332 0 Td
(Tris-HCl)Tj
3.8551 0 Td
(\(pH)Tj
1.8482 0 Td
(7.4\),)Tj
1.9615 0 Td
(1%)Tj
1.4513 0 Td
(Nonidet)Tj
3.5263 0 Td
(P-40,)Tj
2.3074 0 Td
(0.25%)Tj
2.6305 0 Td
(sodium)Tj
3.2315 0 Td
(deoxycho-)Tj
-32.3487 -1.2982 Td
(late,)Tj
1.8481 0 Td
(150)Tj
1.6441 0 Td
(mM)Tj
1.9332 0 Td
(NaCl,)Tj
2.5228 0 Td
(1)Tj
0.6974 0 Td
(mM)Tj
1.9332 0 Td
(EDTA,)Tj
3.0444 0 Td
(1)Tj
0.6973 0 Td
(mM)Tj
1.9332 0 Td
(Na)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 374.2299 282.9543 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 377.8015 284.5984 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(VO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 392.2015 282.9543 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(4)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 395.7732 284.5984 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4478 0 Td
(and)Tj
1.7291 0 Td
(protease)Tj
3.5547 0 Td
(inhibitor)Tj
3.7814 0 Td
(mixture\).)Tj
3.9571 0 Td
(Whole)Tj
2.8913 0 Td
(cell)Tj
-35.9373 -1.2982 Td
(lysates)Tj
2.795 0 Td
(were)Tj
2.1259 0 Td
(centrifuged)Tj
4.7849 0 Td
(at)Tj
0.9524 0 Td
(15,000)Tj
2.8233 0 Td
(g)Tj
0.686 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1735 0 Td
(min)Tj
1.8539 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7517 0 Td
(to)Tj
1.0261 0 Td
(remove)Tj
3.2202 0 Td
(the)Tj
1.4683 0 Td
(cell)Tj
1.5591 0 Td
(debris,)Tj
2.9026 0 Td
(and)Tj
1.7291 0 Td
(superna-)Tj
-33.1877 -1.3039 Td
(tant)Tj
1.7972 0 Td
(protein)Tj
3.1521 0 Td
(was)Tj
1.7007 0 Td
(separated)Tj
4.0082 0 Td
(by)Tj
1.1849 0 Td
(412%)Tj
2.9197 0 Td
(Tris-glycine)Tj
5.0115 0 Td
(precast)Tj
3.05 0 Td
(gel)Tj
1.355 0 Td
(\(KOMA)Tj
3.5546 0 Td
(BIOTECH,)Tj
4.7225 0 Td
(Seoul,)Tj
-32.4564 -1.2983 Td
(Korea\))Tj
2.9707 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(transferred)Tj
4.6374 0 Td
(onto)Tj
2.0806 0 Td
(PVDF)Tj
2.7383 0 Td
(membrane)Tj
4.5636 0 Td
(\(Millipore,)Tj
4.5241 0 Td
(Billerica,)Tj
3.7077 0 Td
(MA\).)Tj
2.3584 0 Td
(Membrane)Tj
4.6431 0 Td
(was)Tj
-35.9713 -1.2983 Td
(blocked)Tj
3.3505 0 Td
(with)Tj
1.9956 0 Td
(5%)Tj
1.4513 0 Td
(non-fat)Tj
3.1975 0 Td
(skim)Tj
2.1657 0 Td
(milk)Tj
2.0466 0 Td
(in)Tj
1.0374 0 Td
(Tris-buffered)Tj
5.5388 0 Td
(saline)Tj
2.5001 0 Td
(including)Tj
4.0422 0 Td
(0.1%)Tj
2.1543 0 Td
(Tween)Tj
2.9084 0 Td
(20)Tj
1.1735 0 Td
(\(TBST\))Tj
-33.5619 -1.2982 Td
(for)Tj
1.3833 0 Td
(1)Tj
0.6973 0 Td
(hour)Tj
2.1543 0 Td
(at)Tj
0.9525 0 Td
(room)Tj
2.4264 0 Td
(temperature.)Tj
5.3518 0 Td
(The)Tj
1.7801 0 Td
(membrane)Tj
4.5695 0 Td
(was)Tj
1.7006 0 Td
(then)Tj
2.0183 0 Td
(incubated)Tj
4.1726 0 Td
(overnight)Tj
4.0875 0 Td
(with)Tj
1.9956 0 Td
(primary)Tj
-33.2898 -1.3039 Td
(ANO1)Tj
2.863 0 Td
(antibody)Tj
3.7644 0 Td
(\(1:500)Tj
2.6872 0 Td
(dilution,)Tj
3.634 0 Td
(ab64085;)Tj
3.7644 0 Td
(Abcam)Tj
3.0896 0 Td
(Inc.,)Tj
1.9843 0 Td
(Cambridge,)Tj
4.9209 0 Td
(MA\).)Tj
2.3641 0 Td
(After)Tj
2.2847 0 Td
(washing)Tj
3.5092 0 Td
(three)Tj
-34.8658 -1.2983 Td
(times)Tj
2.3868 0 Td
(with)Tj
2.0012 0 Td
(TBST,)Tj
2.7213 0 Td
(the)Tj
1.4683 0 Td
(blot)Tj
1.7801 0 Td
(was)Tj
1.7008 0 Td
(further)Tj
3.0274 0 Td
(incubated)Tj
4.1726 0 Td
(for)Tj
1.3888 0 Td
(60)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(at)Tj
0.9525 0 Td
(room)Tj
2.4264 0 Td
(temperature)Tj
5.125 0 Td
(with)Tj
2.0013 0 Td
(an)Tj
1.2018 0 Td
(anti-)Tj
-35.3817 -1.2983 Td
(rabbit)Tj
2.6022 0 Td
(secondary)Tj
4.2746 0 Td
(antibody)Tj
3.7644 0 Td
(\(Santa)Tj
2.7496 0 Td
(Cruz)Tj
2.194 0 Td
(Biotechnology,)Tj
6.2021 0 Td
(CA\).)Tj
2.1316 0 Td
(The)Tj
1.7801 0 Td
(membrane)Tj
4.5695 0 Td
(was)Tj
1.7007 0 Td
(then)Tj
2.0183 0 Td
(washed)Tj
-33.9871 -1.3039 Td
(three)Tj
2.262 0 Td
(times)Tj
2.3925 0 Td
(with)Tj
1.9955 0 Td
(TBST)Tj
2.5002 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(visualized)Tj
4.1385 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
1.4683 0 Td
(ECL)Tj
1.9899 0 Td
(Plus)Tj
1.9162 0 Td
(western)Tj
3.3279 0 Td
(blotting)Tj
3.3618 0 Td
(detection)Tj
3.9345 0 Td
(sys-)Tj
-35.4271 -1.2983 Td
(tem)Tj
1.7575 0 Td
(\(GE)Tj
1.8482 0 Td
(Healthcare,)Tj
4.7961 0 Td
(NJ\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 126.0283 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Quantitative)Tj
5.4992 0 Td
(real-time)Tj
4.0583 0 Td
(PCR)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 109.0204 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Total)Tj
2.313 0 Td
(messenger)Tj
4.422 0 Td
(RNA)Tj
2.2621 0 Td
(\(mRNA\))Tj
3.7587 0 Td
(was)Tj
1.7008 0 Td
(extracted)Tj
3.8834 0 Td
(by)Tj
1.1849 0 Td
(using)Tj
2.3867 0 Td
(a)Tj
0.652 0 Td
(TRIzol)Tj
2.9593 0 Td
(reagent)Tj
3.1862 0 Td
(\(Invitrogen,)Tj
5.0116 0 Td
(Carlsbad,)Tj
-33.7207 -1.3039 Td
(CA\).)Tj
2.1316 0 Td
(Quantitative)Tj
5.2157 0 Td
(Real-time)Tj
4.1045 0 Td
(PCRs)Tj
2.4265 0 Td
(were)Tj
2.126 0 Td
(performed)Tj
4.4673 0 Td
(using)Tj
2.3868 0 Td
(the)Tj
1.474 0 Td
(StepOnePlus)Tj
5.3461 0 Td
(Real-Time)Tj
4.4164 0 Td
(PCR)Tj
-34.0949 -1.2982 Td
(System)Tj
3.05 0 Td
(\(Applied)Tj
3.7474 0 Td
(Biosystems,)Tj
4.8755 0 Td
(CA\))Tj
1.9106 0 Td
(and)Tj
1.7291 0 Td
(Thunderbird)Tj
5.4312 0 Td
(SYBR)Tj
2.5284 0 Td
(qPCR)Tj
2.5682 0 Td
(mix)Tj
1.7745 0 Td
(\(Toyobo,)Tj
3.8778 0 Td
(Osaka,)Tj
2.9196 0 Td
(Japan\).)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 107.1496 23.6976 re
W* n
q
107.0929 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 453.2598 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Downre)Tj
3.3803 0 Td
(gulation)Tj
3.6496 0 Td
(of)Tj
1.0418 0 Td
(ANO1)Tj
2.9126 0 Td
(by)Tj
1.2543 0 Td
(luteolin)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01749)Tj
4.089 0 Td
(35)Tj
0 g
2.0906 0 Td
(March)Tj
2.9622 0 Td
(31,)Tj
1.5874 0 Td
(2017)Tj
34.3562 0 Td
(4)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
58 0 obj
<>stream
endstream
endobj
59 0 obj
<>stream
endstream
endobj
60 0 obj
<>stream
endstream
endobj
61 0 obj
<>stream
endstream
endobj
62 0 obj
<>stream
BT
66.402 0 Td
(13)Tj
ET
Q
endstream
endobj
63 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 10 0 R/Annots 72 0 R/Contents 73 0 R/TrimBox[0 0 612 792]>>
endobj
66 0 obj
<>
endobj
74 0 obj
<>stream
/CIDInit/ProcSet findresource begin
12 dict begin begincmap
/CMapName/Adobe-Identity-UCS def
/CIDSystemInfo<>def
/CMapType 2 def
1 begincodespacerange
<0000>
endcodespacerange
7 beginbfchar
<0010><002D>
<0016><0033>
<0018><0035>
<0024><0041>
<0026><0043>
<002A><0047>
<0037><0054>
endbfchar
endcmap
CMapName currentdict /CMap defineresource pop end end
endstream
endobj
75 0 obj
<>/FontDescriptor 76 0 R/DW 600>>
endobj
76 0 obj
<>
endobj
77 0 obj
<>stream
cvt a &@ fpgm62 h glyf 8
>head 6hhea{<