Fig. 4.

Cell cycle effects. (A) Treatment with reversine induces G₂/M phase accumulation in C2C12 myoblasts. C2C12 myoblasts treated with 20 nM reversine in GM for 48 h were analyzed by FACS based on DNA content (by using PI staining). (B) C2C12 myoblasts were cultured in GM in the presence of 20 nM reversine for 48 h. After removal of reversine (0 h), cells cultured in GM were analyzed by FACS. (C) C2C12 myoblasts synchronized at G₂/M phase and treated with 10 μM U0126 gain multipotency. C2C12 myoblasts synchronized at G₁/S, S, or G₂/M phase were analyzed by FACS. After release from synchronization by the addition of fresh GM in the presence or absence of 10 μM U0126, cells were incubated for 4 h. The compound was then removed, and the cells were cultured in OI or AI conditions for an additional 6 days and analyzed by histocytochemistry. Red, ALP, Oil red O; blue, HT.