Figure 6.

BRD4/CEBPD promotes inflammatory SMC state transition

(A and B) CEBPD gain of function. (C and D) CEBPD loss of function. (E) Treatment of SMCs with TNF-α upregulates BRD4 protein. (F) Effect of pretreatment with bromodomain blocker JQ1. MOVAS cells for stable overexpression of HA-tagged EV or OE-CEBPD were starved in basal medium (DMEM + 0.5% FBS) for 24 h, pretreated with vehicle or JQ1 (1 μM) for 2 h, and then treated with TNF-α (final 20 ng/mL) for 24 h prior to harvest for quantitative real-time PCR (mRNA) or western blot (protein) analysis. For CEBPD silencing, MOVAS cells were transfected with siRNA for 24 h and cultured in fresh starvation medium (0.5% FBS) for another 24 h prior to TNF-α treatment. Quantification: readings from triplicate quantitative real-time PCR reactions were normalized to GAPDH and averaged. The average values from at least 3 independent repeat experiments were then averaged again to calculate mean ± SEM (n = 3–5). Statistics: one-way ANOVA followed by Bonferroni post hoc test; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.