Extended Data Fig. 2: KLF2 regulates BMPER expression.
From: A KLF2-BMPER-Smad1/5 checkpoint regulates high fluid shear stress-mediated artery remodeling
HUVECs were transfected with Ctrl (siCtrl) or KLF2 (siKLF2) siRNA for 4 days, total RNA was extracted and subjected to RNAseq, n = 4 samples for each group. (a) Heatmap showing all differentially expressed genes between siCtrl and siKLF2. (b) Volcano plot of log2 fold change against -log10 p-value (siKLF2 versus siCtrl), red showing differentially up-regulated genes (log2 fold change >1; p-value < 0.05), blue showing differentially down-regulated genes (log2 fold change <−1; p-value < 0.05). Genes related to BMP-Smad1/5 pathway are highlighted. (c) Heatmap show expression of genes relevant to the BMP-Smad1/5 pathway. (d) Q-PCR analysis of BMPER and KLF2 expression, with and without KLF2 KD under indicated FSS levels for 24 h, n = 7 experiments, data are presented as means ± s.e.m. Statistics calculated by two-way ANOVA with Sidak’s multiple comparison tests. ns, not significant. (e) Analysis of KLF4 ChIP-seq dataset from endothelial cells in which constitutively active MEK5 (caMEK5) drove induction of KLF4. caMEK5 triggered increased binding of KLF4 to an intronic region within the BMPER gene, correlating with a near-consensus Klf2/Klf4 motif. The KLF4 binding site correlated with other markers associated with active endothelial enhancer regions, including H3K27Ac and ERG binding.
