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Seung-Bin Yoon, Young-Ho Park, Seon-A Choi, Hae-Jun Yang, Pil-Soo Jeong, Jae-Jin Cha, Sanghoon Lee, Seung Hwan Lee, Jong-Hee Lee, Bo-Woong Sim, Bon-Sang Koo, Sang-Je Park, Youngjeon Lee, Young-Hyun Kim, Jung Joo Hong, Ji-Su Kim, Yeung Bae Jin, Jae-Won Huh, Sang-Rae Lee, Bong-Seok Song, Sun-Uk Kim
Real-time PCR quantification of spliced X-box binding protein 1 (XBP1) using a universal primer method
www.plosone.org
www.plosone.org
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11.9999 0 0 11.9999 200.012 589.096 cm
BT
/F13 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Chemicals)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 572.145 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
0 0 Td
(All)Tj
1.3946 0 Td
(chemicals)Tj
4.1442 0 Td
(and)Tj
1.7348 0 Td
(reagents)Tj
3.5433 0 Td
(used)Tj
2.0579 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(present)Tj
3.1635 0 Td
(study)Tj
2.3924 0 Td
(were)Tj
2.1203 0 Td
(purchased)Tj
4.337 0 Td
(from)Tj
2.2053 0 Td
(Sigma-Aldrich)Tj
-29.5991 -1.3039 Td
(Chemical)Tj
4.0251 0 Td
(Co.)Tj
1.6158 0 Td
(\(St.)Tj
1.559 0 Td
(Louis,)Tj
2.6475 0 Td
(MO,)Tj
2.0807 0 Td
(USA\))Tj
2.4604 0 Td
(unless)Tj
2.6872 0 Td
(otherwise)Tj
4.0819 0 Td
(noted.)Tj
ET
Q
q
1 j
1 J
0.0001 w
11.9999 0 0 11.9999 200.012 531.836 cm
BT
/F13 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Isolation)Tj
3.9354 0 Td
(and)Tj
1.7906 0 Td
(culture)Tj
3.1937 0 Td
(of)Tj
1.0677 0 Td
(various)Tj
3.2882 0 Td
(species)Tj
3.1135 0 Td
(skin)Tj
1.9512 0 Td
(tissues)Tj
2.9764 0 Td
(and)Tj
1.7952 0 Td
(primary)Tj
3.6284 0 Td
(cells)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 514.828 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
0 0 Td
(Primary)Tj
3.4979 0 Td
(fibroblasts)Tj
4.3653 0 Td
(were)Tj
2.1203 0 Td
(isolated)Tj
3.2882 0 Td
(from)Tj
2.2053 0 Td
(ear)Tj
1.4457 0 Td
(skin)Tj
1.8935 0 Td
(and)Tj
1.7348 0 Td
(kidney)Tj
2.9367 0 Td
(in)Tj
1.0374 0 Td
(KSP)Tj
1.9276 0 Td
(mini-pig,)Tj
3.9458 0 Td
(strain)Tj
2.5001 0 Td
(developed)Tj
-32.8986 -1.2982 Td
(by)Tj
1.1792 0 Td
(FARRC)Tj
3.3391 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(])Tj
1.8539 0 Td
(and)Tj
1.7291 0 Td
(cynomolgus)Tj
5.0797 0 Td
(monkey)Tj
3.4696 0 Td
(on)Tj
1.2755 0 Td
(day)Tj
1.6385 0 Td
(1)Tj
0.6916 0 Td
(to)Tj
1.0261 0 Td
(7)Tj
0.6974 0 Td
(after)Tj
2.0352 0 Td
(birth.)Tj
2.4151 0 Td
(Bovine)Tj
2.9991 0 Td
(ear)Tj
1.4513 0 Td
(skin)Tj
1.8935 0 Td
(fibroblasts)Tj
-32.7739 -1.2983 Td
(were)Tj
2.1203 0 Td
(obtained)Tj
3.736 0 Td
(from)Tj
2.2053 0 Td
(local)Tj
2.075 0 Td
(slaughterhouse)Tj
6.2248 0 Td
(cow)Tj
1.8312 0 Td
(ear.)Tj
1.6724 0 Td
(Neonatal)Tj
3.8494 0 Td
(tissues)Tj
2.8177 0 Td
(\(ear)Tj
1.7915 0 Td
(skin)Tj
1.8935 0 Td
(and)Tj
1.7291 0 Td
(kidney\))Tj
3.2768 0 Td
(and)Tj
-35.223 -1.3039 Td
(adult)Tj
2.2506 0 Td
(tissues)Tj
2.8233 0 Td
(\(bovine)Tj
3.2655 0 Td
(ear)Tj
1.4514 0 Td
(skin\))Tj
2.2336 0 Td
(were)Tj
2.1203 0 Td
(chopped)Tj
3.6624 0 Td
(using)Tj
2.3924 0 Td
(surgical)Tj
3.3165 0 Td
(scissors)Tj
3.2315 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(seeded)Tj
2.9083 0 Td
(into)Tj
1.8425 0 Td
(75)Tj
-35.2457 -1.2983 Td
(cm)Tj
ET
Q
q
1 j
1 J
0.0001 w
7.4999 0 0 6.6659 212.428 467.263 cm
BT
/F7 1 Tf
-0.0053 Tc
1 TL
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 218.211 462.841 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
0 0 Td
(T-flask.)Tj
3.2314 0 Td
(C57BL/6N)Tj
4.4957 0 Td
(mouse)Tj
2.8574 0 Td
(embryonic)Tj
4.541 0 Td
(fibroblasts)Tj
4.3654 0 Td
(\(MEFs\))Tj
3.2541 0 Td
(were)Tj
2.1203 0 Td
(isolated)Tj
3.2882 0 Td
(at)Tj
0.9524 0 Td
(day)Tj
1.6328 0 Td
(13.5)Tj
1.8708 0 Td
(\(E13.5\).)Tj
-34.4293 -1.2982 Td
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2.2676 0 Td
(HEK)Tj
2.228 0 Td
(239)Tj
1.6498 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(purchased)Tj
4.3313 0 Td
(from)Tj
2.211 0 Td
(ATCC.)Tj
3.0671 0 Td
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3.4922 0 Td
(cells)Tj
1.9219 0 Td
(and)Tj
1.7291 0 Td
(cell)Tj
1.5591 0 Td
(lines)Tj
2.0692 0 Td
(were)Tj
2.1203 0 Td
(cultured)Tj
3.566 0 Td
(in)Tj
-36.2548 -1.3039 Td
(high-glucose)Tj
5.312 0 Td
(Dulbecco's)Tj
4.5921 0 Td
(Modified)Tj
3.9175 0 Td
(Eagle's)Tj
2.9366 0 Td
(Medium)Tj
3.6737 0 Td
(\(DMEM;)Tj
3.8778 0 Td
(11995,)Tj
2.8233 0 Td
(ThermoFisher\))Tj
6.2985 0 Td
(supple-)Tj
-33.4315 -1.2983 Td
(mented)Tj
3.2597 0 Td
(with)Tj
1.9956 0 Td
(10%)Tj
1.9332 0 Td
(fetal)Tj
1.9049 0 Td
(bovine)Tj
2.931 0 Td
(serum)Tj
2.7213 0 Td
(\(FBS;)Tj
2.3697 0 Td
(16000,)Tj
2.8233 0 Td
(ThermoFisher\))Tj
6.2985 0 Td
(and)Tj
1.7292 0 Td
(1%)Tj
1.457 0 Td
(penicillin/strepto-)Tj
-29.4234 -1.2983 Td
(mycin)Tj
2.7325 0 Td
(\(15140,)Tj
3.1634 0 Td
(ThermoFisher\).)Tj
6.5254 0 Td
(Exceptionally,)Tj
5.845 0 Td
(4)Tj
0.6973 0 Td
(ng/ml)Tj
2.6305 0 Td
(basic)Tj
2.211 0 Td
(fibroblast)Tj
4.0025 0 Td
(growth)Tj
3.0784 0 Td
(factor)Tj
2.5399 0 Td
(\(bFGF;)Tj
-33.4259 -1.3039 Td
(233-FB-025,)Tj
5.1079 0 Td
(R&D)Tj
2.2791 0 Td
(systems\))Tj
3.6396 0 Td
(is)Tj
0.8504 0 Td
(added)Tj
2.6589 0 Td
(to)Tj
1.0261 0 Td
(kidney)Tj
2.9367 0 Td
(tissue)Tj
2.4605 0 Td
(derived)Tj
3.2144 0 Td
(cells.)Tj
ET
Q
q
1 j
1 J
0.0001 w
11.9999 0 0 11.9999 200.012 370.545 cm
BT
/F13 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Oocyte)Tj
3.1653 0 Td
(collection)Tj
4.2757 0 Td
(and)Tj
1.7952 0 Td
(in)Tj
1.063 0 Td
(vitro)Tj
2.2347 0 Td
(maturation)Tj
4.9843 0 Td
(\(IVM\))Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 353.537 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
0 0 Td
(Bovine)Tj
2.999 0 Td
(and)Tj
1.7291 0 Td
(porcine)Tj
3.2825 0 Td
(ovaries)Tj
3.0331 0 Td
(were)Tj
2.1259 0 Td
(obtained)Tj
3.7304 0 Td
(from)Tj
2.2053 0 Td
(a)Tj
0.6577 0 Td
(local)Tj
2.0749 0 Td
(slaughterhouse)Tj
6.2249 0 Td
(and)Tj
1.7291 0 Td
(transported)Tj
4.8755 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
-35.6936 -1.2982 Td
(laboratory)Tj
4.3483 0 Td
(in)Tj
1.0374 0 Td
(0.9%)Tj
2.1544 0 Td
(saline)Tj
2.5001 0 Td
(supplemented)Tj
5.8507 0 Td
(with)Tj
2.0012 0 Td
(75)Tj
/F14 1 Tf
1.1679 0 Td
()Tj
/F7 1 Tf
(g/mL)Tj
2.88 0 Td
(potassium)Tj
4.3143 0 Td
(penicillin)Tj
3.9741 0 Td
(G)Tj
0.9354 0 Td
(and)Tj
1.7348 0 Td
(50)Tj
/F14 1 Tf
1.1679 0 Td
()Tj
/F7 1 Tf
(g/ml)Tj
-34.0665 -1.2983 Td
(streptomycin)Tj
5.5105 0 Td
(sulfate)Tj
2.7949 0 Td
(and)Tj
1.7348 0 Td
(maintained)Tj
4.7849 0 Td
(at)Tj
0.9524 0 Td
(2530C.)Tj
3.9231 0 Td
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1.338 0 Td
(recovered)Tj
4.1442 0 Td
(mouse)Tj
2.8573 0 Td
(embryos,)Tj
3.8891 0 Td
(C57BL/6J)Tj
-31.9292 -1.3039 Td
(female)Tj
2.8516 0 Td
(mice)Tj
2.1543 0 Td
(were)Tj
2.126 0 Td
(super-ovulated)Tj
6.1908 0 Td
(with)Tj
1.9956 0 Td
(5)Tj
0.6973 0 Td
(IU)Tj
1.2926 0 Td
(of)Tj
1.0148 0 Td
(pregnant)Tj
3.8267 0 Td
(mare)Tj
2.2677 0 Td
(serum)Tj
2.7269 0 Td
(gonadotropin)Tj
5.7203 0 Td
(\(PMSG;)Tj
-32.8646 -1.2983 Td
(Sigma-Aldrich\),)Tj
6.6443 0 Td
(followed)Tj
3.651 0 Td
(by)Tj
1.1792 0 Td
(5)Tj
0.6973 0 Td
(IU)Tj
1.2926 0 Td
(of)Tj
1.0148 0 Td
(human)Tj
3.0784 0 Td
(chorionic)Tj
4.0649 0 Td
(gonadotropin)Tj
5.7203 0 Td
(\(hCG;)Tj
2.6929 0 Td
(Sigma-)Tj
3.0273 0 Td
(Aldrich\))Tj
-33.063 -1.2982 Td
(48)Tj
1.1678 0 Td
(h)Tj
0.7484 0 Td
(apart)Tj
2.279 0 Td
(and)Tj
1.7291 0 Td
(mated)Tj
2.7213 0 Td
(with)Tj
1.9955 0 Td
(male)Tj
2.143 0 Td
(C57BL/6J)Tj
4.0876 0 Td
(mice.)Tj
2.381 0 Td
(Additional)Tj
4.5298 0 Td
(details)Tj
2.7779 0 Td
(were)Tj
2.1203 0 Td
(performed)Tj
4.4674 0 Td
(as)Tj
1.0148 0 Td
(previ-)Tj
-34.1629 -1.2983 Td
(ously)Tj
2.313 0 Td
(described)Tj
4.0535 0 Td
([)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7405 0 Td
(19)Tj
0 g
(].)Tj
ET
Q
q
1 j
1 J
0.0001 w
11.9999 0 0 11.9999 200.012 248.258 cm
BT
/F13 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Universal)Tj
4.2189 0 Td
(primer)Tj
3.1323 0 Td
(design)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 231.307 cm
BT
/F7 1 Tf
-0.01 Tc
1 TL
0 0 Td
(In)Tj
1.0941 0 Td
(order)Tj
2.3924 0 Td
(to)Tj
1.0148 0 Td
(effectively)Tj
4.1443 0 Td
(detect)Tj
2.5795 0 Td
(the)Tj
1.4513 0 Td
(ER)Tj
1.4003 0 Td
(stress)Tj
2.3584 0 Td
(in)Tj
1.0262 0 Td
(various)Tj
3.0954 0 Td
(animal)Tj
2.9366 0 Td
(species,)Tj
3.1748 0 Td
(u/s/tXBP1)Tj
4.2803 0 Td
(primer)Tj
2.9537 0 Td
(were)Tj
-33.9021 -1.3039 Td
(designed)Tj
3.7133 0 Td
(to)Tj
1.0148 0 Td
(relative)Tj
3.0898 0 Td
(the)Tj
1.4513 0 Td
(26)Tj
1.1622 0 Td
(bp)Tj
1.2302 0 Td
(of)Tj
1.0035 0 Td
(the)Tj
1.4513 0 Td
(XBP1)Tj
2.4661 0 Td
(removed)Tj
3.702 0 Td
(by)Tj
1.1736 0 Td
(IRE1)Tj
2.1997 0 Td
(gene.)Tj
2.279 0 Td
(Primer)Tj
2.9934 0 Td
(was)Tj
1.6837 0 Td
(designed)Tj
3.7134 0 Td
(by)Tj
1.1735 0 Td
(com-)Tj
-35.5008 -1.2982 Td
(paring)Tj
2.7836 0 Td
(the)Tj
1.457 0 Td
(sequence)Tj
3.8097 0 Td
(of)Tj
1.0035 0 Td
(human,)Tj
3.2654 0 Td
(monkey,)Tj
3.651 0 Td
(cow,)Tj
2.0353 0 Td
(pig)Tj
1.4513 0 Td
(and)Tj
1.7122 0 Td
(mouse,)Tj
3.05 0 Td
(using)Tj
2.3584 0 Td
(of)Tj
1.0035 0 Td
(the)Tj
1.4513 0 Td
(base)Tj
1.9219 0 Td
(sequence)Tj
3.8097 0 Td
(that)Tj
-34.7638 -1.2983 Td
(are)Tj
1.4286 0 Td
(well)Tj
1.7972 0 Td
(conserved)Tj
4.2065 0 Td
(XBP1)Tj
2.4662 0 Td
(gene.)Tj
2.2733 0 Td
(For)Tj
1.6101 0 Td
(the)Tj
1.4513 0 Td
(normalization)Tj
5.8054 0 Td
(on)Tj
1.2585 0 Td
(the)Tj
1.457 0 Td
(gene)Tj
2.0523 0 Td
(expression,)Tj
4.6261 0 Td
(a)Tj
0.6463 0 Td
(primer)Tj
2.9594 0 Td
(was)Tj
1.6781 0 Td
(pro-)Tj
-35.7163 -1.3039 Td
(duced)Tj
2.6191 0 Td
(for)Tj
1.3663 0 Td
(the)Tj
1.4514 0 Td
(GAPDH,)Tj
3.8721 0 Td
(HPRT,)Tj
2.9763 0 Td
(H2afz,)Tj
2.7893 0 Td
(18S)Tj
1.6271 0 Td
(rRNA,)Tj
2.8289 0 Td
(ActB)Tj
2.1827 0 Td
(genes)Tj
2.4094 0 Td
(known)Tj
2.9764 0 Td
(as)Tj
1.0035 0 Td
(housekeeping)Tj
5.6352 0 Td
(genes,)Tj
-33.7377 -1.2983 Td
(and)Tj
1.7121 0 Td
(a)Tj
0.6463 0 Td
(total)Tj
1.9729 0 Td
(amount)Tj
3.3221 0 Td
(of)Tj
1.0035 0 Td
(XBP1)Tj
2.4605 0 Td
(gene)Tj
2.0579 0 Td
(expression)Tj
4.405 0 Td
(was)Tj
1.6838 0 Td
(used)Tj
2.0352 0 Td
(to)Tj
1.0092 0 Td
(the)Tj
1.457 0 Td
(precise)Tj
2.965 0 Td
(ER)Tj
1.3946 0 Td
(stress.)Tj
2.5795 0 Td
(\()Tj
0.83 0.64 0.02 0 k
(S1)Tj
1.4967 0 Td
(Table)Tj
0 g
(\))Tj
ET
Q
q
1 j
1 J
0.0001 w
11.9999 0 0 11.9999 200.012 139.011 cm
BT
/F13 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Stimulation)Tj
5.1779 0 Td
(of)Tj
1.0678 0 Td
(ER)Tj
1.4504 0 Td
(stress)Tj
2.5417 0 Td
(in)Tj
1.0678 0 Td
(various)Tj
3.2882 0 Td
(cells)Tj
2.0126 0 Td
(and)Tj
1.7905 0 Td
(embryos)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 122.003 cm
BT
/F7 1 Tf
-0.018 Tc
1 TL
0 0 Td
(ER)Tj
1.3832 0 Td
(stress)Tj
2.3131 0 Td
(was)Tj
1.6554 0 Td
(induced)Tj
3.3676 0 Td
(in)Tj
1.0091 0 Td
(various)Tj
3.0444 0 Td
(kinds)Tj
2.3471 0 Td
(of)Tj
0.9864 0 Td
(somatic)Tj
3.2372 0 Td
(cells)Tj
1.8481 0 Td
(and)Tj
1.6895 0 Td
(embryos)Tj
3.5659 0 Td
(used)Tj
2.0013 0 Td
(in)Tj
1.0091 0 Td
(the)Tj
1.4287 0 Td
(experiment)Tj
4.6374 0 Td
(that)Tj
-35.5235 -1.2982 Td
(were)Tj
2.0636 0 Td
(exposed)Tj
3.3505 0 Td
(to)Tj
0.9978 0 Td
(tunicamycin)Tj
5.074 0 Td
(\(TM,)Tj
2.2336 0 Td
(2)Tj
/F14 1 Tf
0.686 0 Td
()Tj
/F7 1 Tf
(g/mL\))Tj
3.1351 0 Td
(or)Tj
1.0715 0 Td
(thapsigargin)Tj
5.006 0 Td
(\(TG,)Tj
2.0579 0 Td
(50)Tj
1.1395 0 Td
(nM\))Tj
1.9616 0 Td
(for)Tj
1.3436 0 Td
(various)Tj
3.0444 0 Td
(time)Tj
1.9729 0 Td
(peri-)Tj
-35.138 -1.3039 Td
(ods.)Tj
1.7858 0 Td
(The)Tj
1.7404 0 Td
(chemicals)Tj
4.0195 0 Td
(were)Tj
2.0636 0 Td
(provided)Tj
3.7077 0 Td
(as)Tj
0.9865 0 Td
(lyophilized)Tj
4.4617 0 Td
(powders)Tj
3.5149 0 Td
(and)Tj
1.6895 0 Td
(dissolved)Tj
3.77 0 Td
(in)Tj
1.0035 0 Td
(DMSO)Tj
3.016 0 Td
(\(TM,)Tj
2.2337 0 Td
(TG\))Tj
1.8482 0 Td
(to)Tj
-35.841 -1.2983 Td
(give)Tj
1.7744 0 Td
(stock)Tj
2.2337 0 Td
(solutions.)Tj
3.9288 0 Td
(The)Tj
1.7405 0 Td
(respective)Tj
4.0478 0 Td
(diluents)Tj
3.3052 0 Td
(were)Tj
2.0693 0 Td
(used)Tj
2.0012 0 Td
(as)Tj
0.9865 0 Td
(a)Tj
0.6406 0 Td
(negative)Tj
3.4242 0 Td
(control)Tj
3.0217 0 Td
(in)Tj
1.0092 0 Td
(each)Tj
1.9729 0 Td
(experiment.)Tj
ET
Q
q
1 j
1 J
0.0001 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
Q
q
36 743.357 94.4504 15.5339 re
W* n
q
0.148 0 0 0.147998 36 743.357 cm
q
/I4 Do
Q
Q
Q
q
0 0 612 792 re
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