H3K9me3 levels on myogenic genes increased in C2C12 myoblasts after depleting HP1α.

<p><i>A.</i> Schematic diagram of the genomic structure of the mouse Lbx1 gene and locations of primers used for subsequent ChIP experiments. <i>B.</i> Protein-DNA complexes from cross-linked chromatin extracted from C2C12 myoblasts cultured in GM were immunoprecipitated with HP1α or mouse IgG. Bound DNA was amplified using the indicated PCR primers. <i>C, D,</i> C2C12 myoblasts were transfected with indicated siRNA, 48 hours after transfection, cross-linked chromatin was extracted and immunoprecipitated with indicted antibodies. <i>Lbx1</i> exon 2 (C) or <i>Lbx1</i> genomic sequences including exon 1, intron and exon 2 (D) were amplified. E. C2C12 myoblasts were transfected with the indicated siRNA, 48 hours after transfection total cell lysates were subjected to Western blotting with the indicated antibodies. <i>F.</i> C2C12 myoblasts were transfected with indicated siRNA, 48 hours after transfection cross-linked chromatin was extracted and immunoprecipitated with anti-H3K9me3 antibody. Precipitated DNA was used for PCR with primers spanning the MEF2-binding site on the myogenin gene promoter.