Figure 4
From: Hypoxia upregulates expression of human endosialin gene via hypoxia-inducible factor 2

The binding of hypoxia-inducible factor (HIF) subunits to hypoxia-responsive elements in the endosialin promoter. Chromatin immunoprecipitation assays using FIB-3 cells treated with 100 μ M DIP for 4 and 24 h, respectively. Untreated cells were used as a negative control. The cells were incubated with formaldehyde for crosslinking, DNA was extracted and immunoprecipitated with antibodies specific for HIF subunits or for human IgG antibodies to visualise background signal. Recovered DNA was used in PCR with primers flanking respective HREs. DNA samples from non-immunoprecipitated extracts were amplified as input. HIF-2α was found to bind to HRE −976/−969, but not to the other HREs.