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. Author manuscript; available in PMC: 2014 Sep 3.
Published in final edited form as: Nat Struct Mol Biol. 2007 Nov 18;14(12):1150–1156. doi: 10.1038/nsmb1316

Figure 4.

Figure 4

Inhibition of p38 MAPK activity prevents recruitment of Ash2L to the Myog promoter. (a) Protein extracts prepared from C2C12 cells treated with SB203580 (+SB) or untreated (−SB) under either growth (0 h) or differentiation (48 h) conditions were analyzed by western blotting with indicated antibodies. (b) Inhibition of p38 kinase activity blocks transcription of several muscle-specific genes in C2C12 cells. RNA was isolated from +SB or −SB C2C12 cells under either growth or differentiation conditions. After reverse transcription, random primed cDNA was subjected to semiquantitative PCR analysis with primers specific for the genes indicated. (c) Inhibition of p38 kinase activity blocks recruitment of Ash2L and prevents H3K4me3 modification at the Myog promoter. ChIP was used to measure relative enrichment of the indicated proteins at the Myog promoter in C2C12 cells differentiated (48 h) in the presence or absence of SB203580. After deproteination, immunopurified DNA was quantified by qRT-PCR with hydrolysis probes. Average values of duplicate qRT-PCR reactions are shown; error bars represent s.d. Each experiment was performed at least twice with independent chromatin samples and yielded similar results both times.