ChIP assays were carried out as previously described using 150 µg of chromatin for EZH2 and MyoD, and 50 µg of chromatin for modified histones. In brief, chromatins were immunoprecipitated with anti-MyoD antibody ([sc-760] Santa Cruz Biotechnology), anti-EZH2 antibody (39901, Active Motif), anti trimethyl-histone H3K27 (07-449, Millipore), anti trimethyl-histone H3K4 (07-473, Millipore), normal rabbit (Millipore, 12-370) or normal mouse IgG (Millipore, 12-371). qPCR analysis was performed in triplicate for each of three independent experiments, using 5 ng of DNA, Go Taq qPCR Master Mix (Promega) and the following primer pairs:p57 prom Maternal p57i Paternal p57i β-globin prom β-Actin prom Amylase prom Albumin prom Timm prom p57i
F: 5′5′ACTGAGAGC-AAGCGAACAGG-3′
R: 5′ACCTGGCTGATTGGTGATGG-3′;
F: 5′-CAGATCTGACCTCAGACCCA-3′
R: 5′-CCTGTTCCTCGCCGTCCC-3′;
F: 5′CAGATCTGACCTCAGACCCG-3′
R: 5′-GACCTGTTCCTCGCCATCCT-3′;
F: 5′-GAAGCCTGATTCCGTAGAGC-3′
R: 5′-CAACTGATCCTACCTCACCTTATATGC-3′;
F: 5′-GTGACATCCACACCCAGAGG-3′
R: 5′-GAATAGCCTCCGCCCTTG-3′;
F: 5′-TCAGATGGGAGGACTGCTATTGT-3′
R: 5′-GCTCACATTCCTTGGCAATATCA-3′;
F: 5′-AATCGTCTTTGAGGCACCAG-3′
R: 5′-GCTCAATCTTCCCAAACAGG-3′;
F: 5′-ACGGATGTGGCCCTTCTGGCT-3′
R: 5′-CCGCTGCGAAACGCCCACAA-3′;
F: 5′-AACTTCCAGCAGGATGTGCC-3′
R: 5′-CATCCACTGCAGACGACCAG-3′