A. BRD4 silencing reduces DOT1L expression. Mouse SMCs (MOVAS) were transduced with lentivirus to express scrambled or gene-specific shRNAs. The transduced cells were allowed to recover in fresh medium for 24h and then starved overnight in basal medium prior to stimulation with PDGF-BB (20ng/ml) for 24 h. Immunoblots: Mean ± SEM, n = 3 independent repeat experiments. qRT-PCR: Mean ± SD, n = 3 replicates. ANOVA and Tukey test: *p<0.05, compared to any of the other 3 bars.
B. BRD4 silencing down-regulates AURKB. Rat primary aortic SMCs were transfected with scrambled or BRD4-specific siRNA, and stimulated with PDGF-BB (20ng/ml, 48h). Mean ± SEM, n= 3 independent repeat experiments. Student’s t-test: *p<0.05.
C. BRD4 and H3K27ac co-occupy the genomic region of Aurkb. The data was retrieved by using accession number GSE194390 (see our recent report45). Integrative genomics viewer (IGV) tracks show the comparison of normalized ChIP-seq peaks between injured (+, light color) and uninjured sham-control (-, dark color) arteries. The box on the left highlights the Aurkb TSS region where the binding of H3K27ac and BRD4 increased after arterial injury. Non-specific input indicates low background noise.