Role of histone acetylation in SRF signaling.

<p>(A) SRF transcriptional activity in human keratinocytes expressing the SRF luciferase reporter and TK renilla control. Cells were treated overnight with or without 2 µM SB202190 or 200 nM TSA, then stimulated for 7 h with 10% FBS. Data are means ± SEM of 4 replicates from one experiment (representative of 3 independent experiments). *P<0.05 for effect of SB202190, +P<0.05 for effect of TSA. (B) Expression of endogenous SRF target genes measured by real-time RT-PCR. Cells were serum-starved overnight with or without 200 nM TSA, then stimulated with 10% FBS-containing medium for 1 h. Data are means ± SEM of 3 experiments. *P<0.05 compared to carrier. (C) Western blot of total and acetylated histone H3 in cells after 24 h treatment with carrier, 1 µM cytochalasin D, 2 µM SB202190, or 200 nM TSA. (D) ChIP of acetylated H3 in cells following 24 h treatment with 2 µM SB202190. DNA sequence flanking the SRE regulatory elements in the <i>FOS</i> and <i>JUNB</i> genes were detected by real-time RT-PCR, and the centromeric region of chromosome 11 (<i>SATCEN11</i>) was used as a negative control. Enrichment from the anti-AcH3 IP is reported relative to anti-FLAG control.