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. 2012 Dec 15;23(24):4778–4785. doi: 10.1091/mbc.E12-04-0311

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© 2012 Ling et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — MyoD methylation is relevant in the inhibition of myogenesis by Sharp-1. (A) C2C12 cells overexpressing Sharp-1 were transfected with MyoD and MyoD(K104R). MyoD expression was analyzed by Western blot. (B-D) Control (vector), MyoD, and MyoD(K104R) cells were analyzed for differentiation with anti-MHC antibody (B), myogenic index (C), and troponin T expression by Western blot (D). (E) C3H10T1/2 cells were transfected with MyoD, MyoD(K104R), Myc-Sharp-1, and FLAG-G9a. MyoD was immunoprecipitated and probed with anti-Me Lys antibody. Expression of Sharp-1, MyoD, and G9a was analyzed in lysates. (F) ChIP assays were performed in Sharp-1–overexpressing C2C12 cells transfected with MyoD and MyoD(K104R). H3K9me2 was analyzed by ChIP assays on the myogenin promoter at D2 of differentiation. Error bars indicate mean ± SD.