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Hypoxic activation of HIF-1α directly regulates the transcriptional activity of ERRγ.

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posted on 19.02.2013, 22:46 by Ja Hee Lee, Eun-Jin Kim, Don-Kyu Kim, Ji-Min Lee, Seung Bum Park, In-Kyu Lee, Robert A. Harris, Mi-Ock Lee, Hueng-Sik Choi

(A) HepG2 cells were transfected with hERRγ-Luc. After 24 h of the transfection, HepG2 cells were exposed to hypoxia for indicated time period. Experiments were carried out in triplicate and data are expressed as the fold activation relative to the control. (B–C) HepG2 cells were transfected with hERRγ (−2 kb)-Luc, hERRγ (−1 kb)-Luc, hERRγ (−0.5 kb)-Luc, hERRγ (−0.3 kb)-Luc, hERRγ (HREmt1)-Luc, hERRγ (HREmt2)-Luc, hERRγ (HREmt1+2)-Luc. After 24 h of the transfection, HepG2 cells were exposed to hypoxia for 9 hr and analyzed using luciferase and β-galactosidase assay. Experiments were performed in duplicate and data are expressed as the fold activation relative to the control. (D) ChIP assay: HepG2 cell was exposed to hypoxia for 9 hr. Input represents 10% of purified DNA in each sample. Cell extracts were immunoprecipitated with anti-HIF-1α and purified DNA samples were employed for Q-PCR with primers binding to HRE1 (−1080 to −849) and HRE2 (−508 to −295) and distal site (−1826 to −1586) on the ERRγ gene promoter. All data are representative of at least three independent experiments. Error bars show ± S.E.M. ***P<0.001 by two-tailed Student t-test.

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