Fig. 4: Targeting BET protein suppresses key EC activities under normal growth conditions.
From: Three-dimensional CRISPR screening reveals epigenetic interaction with anti-angiogenic therapy
a Effects of siRNA-mediated knockdown of BRD2, BRD3, or BRD4 on survival and/or proliferation of ECs. Error bars represent ± SEM (n = 2 independent experiments). b Effects of I-BET762 (1000 nM) or JQ1 (300 nM) on survival and/or proliferation of ECs. Error bars represent ± SEM (n = 2 independent experiments for all conditions except n = 4 independent experiments for DMSO). c Effects of BETi on nuclear DNA content of ECs. Nuclei were released and analyzed on day 3 of BETi treatment. Error bars represent ± SEM (n = 2 independent experiments for all conditions except n = 3 independent experiments for DMSO). *, adjusted P value < 0.05; **, adjusted P value < 0.01; ***, adjusted P value < 0.001 (versus DMSO, one-way ANOVA with Dunnett’s multiple comparison test performed). d Scratch wound migration assay analysis of the effect of I-BET762 (1000 nM) or JQ1 (300 nM) on EC migration. For imaging, ECs were stained using CellTracker Green CMFDA dye at 0 h (T0) then fixed and stained with phalloidin CF488A and Hoechst 33342 at 24 h after scratching (T24). Images were assembled as collages. Scale bars, 200 μm. e Quantification of EC migration. f Quantification of cell number at T24 normalized to T0. Error bars represent ± SEM (n = 3 independent experiments). ***P value < 0.001 (two-tailed unpaired t test).
