1. HIF1 binds and stimulates the VEGFA promoter in normoxia.
HIF1 binds and stimulates the VEGFA promoter in normoxia. (A) Schematic presentation of a human VEGFA promoter, −1312/−140 (1173, bp) corresponds to the region identified by ChIP sequencing (Figure 3C). Potential E2F and HIF binding sites (BS), identified by Matinspector software, are indicated. Positioning is based on the transcriptional start site (bent arrow). Two promoter elements (indicated as element 1 or 2) contain closely located E2F‐ and HIF‐BS. Sites indicated with asterisks are 100% identical between human and mouse. (B) HIF1α ChIP (VEGFA promoter) performed on normoxic HeLa cells. (C) Upper panel, activity of a 47‐bp (−985/−939) VEGFA reporter plasmid, containing the −974 HIF‐BS, co‐transfected with HIF1α or scrambled (scr) siRNA in normoxic U2OS cells. Lower panels, western blot for HIF1α and loading control HDAC1. (D) U2OS cells transfected with HIF1α or scr siRNA, grown under normoxic (black bars) or hypoxic conditions (white bars). Upper panel shows VEGFA mRNA levels and lower panel shows western blot for HIFα and HDAC1 (loading). (E) Similar to (D) but now for HeLa cells. 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:
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