Figure 3

BET action on proliferative myoblasts. A. Simplified design of experimental procedures: myoblasts were grown until 40% confluence in GM and then the medium was switched in GM with 10 mM BET or in DM. The experiment continued until the cells reached subconfluence. B. Growth curve trend: 10 mM BET did not influence C2C12 proliferative potential. C. Cell cycle gene expression after 24 h of treatment: the gene expression profile in the three experimental conditions was similar. D. Representative blot of analyzed proteins are shown. E-F. Western blot analysis: BET 10 mM significantly improved MyoD amount, but did not modified p21 content. G-H. MyoD immunofluorescence and phase contrast microscopy images revealed that BET 10 mM myoblasts started to assume an elongated shape, similarly to DM. We performed three independent experiments. For gene expression analysis data as presented in as fold-change (ΔΔCt) of threshold cycle (Ct) mean ± SD. Significance: d = p ≤ 0.02, e = p ≤ 0.01. Scale bar: 200 μm.