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. 2019 Apr 30;10:933. doi: 10.3389/fimmu.2019.00933

Figure 3.

Figure 3

I-BET151 has no effect on MAPK and NFκB activation in gingival fibroblasts (GFs) and new protein synthesis is not required for inflammatory gene suppression by I-BET151. (A) Western blot analysis of phospho (p)-ERK, p-p38, IκBα, p-p65(Ser536), and tubulin (Tub) in total cell lysates of GFs after 30 min treatment with DMSO or 1 μM I-BET151 followed by stimulation with 10 ng/ml TNF for 5, 20, or 60 min. (B) Western blot analysis of total acetylated lysine (AcLys), acetyl-histone H3(Lys18) (AcH3), acetyl-H4(Lys8) (AcH4), and total H3 in cell lysates of GFs treated with DMSO or 1 μM I-BET151 for 30 min prior to TNF (10 ng/ml) stimulation for 4 or 24 h. Cells treated with SAHA for 4 h were used as a positive control for lysine hyperacetylation. Data representative of 2–3 independent experiments are shown in (A,B). (C) Relative mRNA expression of IL1B, CCL2, and COX2 in GFs treated with DMSO or 1 μM I-BET151 in the presence or absence of cycloheximide (CHX) for 30 min prior to simulation with 10 ng/ml TNF for 4 h analyzed by qPCR (mean + SEM; n = 4; % of suppression compared to DMSO control are depicted in each graph).