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Figure 3 | Epigenetics & Chromatin

Figure 3

From: Chromatin regulated interchange between polycomb repressive complex 2 (PRC2)-Ezh2 and PRC2-Ezh1 complexes controls myogenin activation in skeletal muscle cells

Figure 3

Msk1-dependent H3S28 phosphorylation affects PRC2-Ezh2 chromatin displacement from muscle regulatory regions. Chromatin immunoprecipitation (ChIP) analysis was performed on chromatin prepared from C2C12 cells cultured in growth medium (GM) or differentiation medium (DM) 48 h after induction of differentiation with or without Msk1 inhibitor H89 (5 μM) using histone H3 phosphorylation at serine 28 (H3S28ph), acetylated histone 3 (AcH3), Msk1, Ezh2 and H3K27me3 antibodies. The precipitated DNA fragments were subjected to real-time PCR analysis with primers amplifying the myogenin (MyoG) promoter (A), muscle creatine kinase (mCK) enhancer (B) and mCK promoter (C). ChIP values are presented as relative enrichments to myoblasts. Levels of H3S28ph, AcH3 and H3K27me3 were normalised to histone H3 density. Data represent the average of three independent experiments. Error bars represent SD. (D) Recombinant Msk1 was incubated with a histone H3 (residues 21-33) peptide either unmodified or modified with the K27me3 or S28ph, and a kinase assay was performed (cpm = counts per min).

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Epigenetics & Chromatin

ISSN: 1756-8935