Promoter contact with dynamic H3K27ac loci in H1 and H4-12 clusters was stimulated by VEGFA and required EP300 activity. (A–D) Chromatin confirmation capture of dynamic H3K27ac sites (red lines) near DUSP5, NR4A1, KDR, and CD34. (Black boxes) Regions of H3K27ac enrichment; chromatin looping was attenuated by pretreatment of HUVEC cells with C646 for 30 min. (Red lines) Dynamic H3K27ac sites. (Gray bars) Promoter anchor primers and fragments graphed to the right. (Vertical black lines) Cutting sites of restriction enzyme used for chromatin conformation capture. (*) P < 0.05. (E) EP300 acetyltransferase activity was required for activation of gene expression downstream from VEGFA. Gene expression was measured by qRT-PCR during VEGFA stimulation in the presence or absence of the EP300 inhibitor C646.
