Figure 2

Myogenin (MyoG) and muscle creatine kinase (mCK) muscle markers are differentially bound by PRC2-Ezh2 and PRC2-Ezh1 complexes in C2C12 cell lines. (A) (I) Expression levels of MyoG were measured by real-time PCR in myoblasts grown in growth medium (GM) or differentiation medium (DM), 2, 4 or 8 days after induction of differentiation. The transcription levels were normalised to Gapdh expression and represent the mean of three independent experiments ± SD. Fold enrichment was calculated in comparison to myoblasts in GM. (II, III) Chromatin immunoprecipitation (ChIP) analysis of chromatin prepared from cells cultured in GM or in DM for 2, 4 and 8 days with Ezh2, Suz12, Ezh1 (II) and RNA polymerase II (RNA Pol II) (III) antibodies. The precipitated DNA fragments were subjected to real-time PCR analysis using primers designed within MyoG promoter. ChIP enrichments are presented as a percentage (%) of the input. Data are shown as an average of three independent experiments ± SD. (B) (I) Expression levels of mCK were measured by real-time PCR in myoblasts grown in GM or DM, 2, 4, or 8 days after induction of differentiation. The transcription levels were normalised to Gapdh expression and represent the mean of three independent experiments ± SD. Fold enrichment was calculated in comparison to myoblasts in GM. (II, III) ChIP analysis of chromatin prepared from cells cultured in GM or in DM for 2, 4 and 8 days with Ezh2, Suz12, Ezh1 (II) and RNA Pol II (III) antibodies. The precipitated DNA fragments were subjected to real-time PCR analysis using primers designed within mCK enhancer. ChIP enrichments are presented as percentage of the input. Data are shown as average of three independent experiments ± SD.