Figure 6.

ERG (ETS-related gene) contributes to enhancer activation in human umbilical vein endothelial cell (HUVEC). A, Volcano plot showing log₂ fold change (FC) vs –log (P value) of differential H3K27ac enrichment at siCtl H3K27ac regions in response to ERG knockdown. Loss and gain enhancer regions are selected by −1≥log₂FC≥1; −log (P value)≥4. B, Heatmap of H3K27ac enrichment over input in all loss and gain regions. Signal ±5 kb from the center of H3K27ac siCtl or siERG regions as tag density/bp. C, Chromatin immunoprecipitation-quantitative polymerase chain reaction (ChIP-qPCR) of H3K27ac at enhancers of CLDN5 (claudin-5), DLL4 (Delta-like protein 4), and VWF (von Willebrand factor) with a negative control gene desert, in siCtl vs siERG-treated HUVEC. Graph represents fold change over IgG, n=3. *P<0.05, paired 2-tailed t test. D, Boxplot representing log₂FC from transcriptome profiling data following ERG knockdown in H3K27ac regions changed in response to siERG (as defined in A). P<0.0001, Kruskal-Wallis test and post hoc test using Wilcoxon rank-sum test, ****P<0.0001; *P<0.05. E, Pathway analysis of genes associated with loss and gain of H3K27ac. F, Boxplot showing the log₂FC of MED (Mediator complex subunit)-1 occupancy in siERG-treated HUVEC in H3K27ac regions changed in response to siERG. P<0.0001, Kruskal-Wallis test and post hoc test using Wilcoxon rank-sum test, ****P<0.0001. TGF indicates transforming growth factor. BMP indicates bone morphogenetic protein; HIF, hypoxia inducible factor; TGF, transforming growth factor; and VEGFR, vascular endothelial growth factor receptor.