E2F7/8 and HIF1 are required for hypoxic VEGFA expression.

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E2F7/8 and HIF1 are required for hypoxic VEGFA expression. E2F7/8 and HIF1 are required for hypoxic VEGFA expression. (A) U2OS cells were transfected with a total of 150 pmol siRNA: 75 pmol E2F7 plus HIF1α, or 75 pmol E2F7 or HIF1α plus 75 pmol scr. Transfected cells were cultured in normoxia or hypoxia. qPCR was performed for indicated transcripts. Lower two panels show HIF1α and HDAC1 (loading) protein levels. (B) Doxycycline‐inducible E2F7‐EGFP or control doxycycline‐inducible EGFP cells were treated with 20 h dox, 4 h hypoxia, 20 h dox of which the last 4 h in combination with hypoxia, or left untreated. Upper panel shows qPCR analysis of VEGFA mRNA. Lower panels display western blot for GFP, HIF1α or ERK (loading). (C) E2F7 and HIF1α ChIPs were performed on hypoxic HeLa cells. ChIP on the E2F1 promoter served as a positive control. A non‐specific region 700 basepairs upstream of the E2F binding site in the E2F1 promoter served as a negative control. All quantified data are presented as the average (±s.d.) compared to the control condition in three independent experiments. Bart G M W Weijts et al. EMBO J. 2012;31:3871-3884 © as stated in the article, figure or figure legend