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. 2014 Sep 26;24(10):1231–1249. doi: 10.1038/cr.2014.127

Figure 7.

Figure 7

SUV39H1 methylates Runx2 at Lys245. (A) HEK293T cells were co-transfected with Myc-Runx2 and Flag-SUV39H1, Flag-G9a or Flag-CBFβ plasmids. Cells were subjected to immunoprecipitation with anti-Myc or anti-Flag, and coprecipitated proteins were immunoblotted. Specific signals for precipitated proteins are asterisked. (B) The differentiation of C2C12 cells was induced by BMP2 treatment for 24 h. Cell lysates were subjected to immunoprecipitation with anti-SUV39H1, and the precipitated proteins were immunoblotted with anti-Runx2. (C) Recombinant GST-R2 peptide was incubated with S-adenosylmethionin (SAM), Flag-SUV39H1 and/or Flag-SBP-PHF2 (F/S-PHF2) peptide at 4 oC or 37 oC for 60 min. The methylation of GST-R2 peptide was detected using anti-methylated lysine antibody (upper panel), and proteins in the reaction mixtures were immunoblotted (bottom panel). (D) HEK293T cells were co-transfected as indicated and the cell lysates were subjected to immunoprecipitation with an anti-Lys-me antibody. Precipitated F/S-R2 or K245A mutants were identified using anti-Flag.