FIGURE 7.

HIF-1α binds to the FBLN5 promoter in hypoxic cells. The relative in vivo association of HIF-1α with the human FBLN5 promoter was analyzed by ChIP in HUVEC incubated under normoxic or hypoxic conditions. Chromatin was sheared by sonication, and immunoprecipitations were performed with an anti-HIF-1α antibody or a nonspecific goat IgG. A, the enrichment of HIF-1α was quantified by real-time PCR using FBLN5 promoter-specific primers. PCR reactions using total input DNA saved before immunoprecipitation and H₂O were also carried out. Data were normalized to the total input DNA and represented as mean ± S.E. of two independent experiments performed in duplicate (*, p < 0.05 versus control cells). B, agarose gel electrophoresis of PCR products amplified using specific primers for both FBLN5 and VEGF promoters. Equal input DNA and control IgG immunoprecipitations are shown.