FIGURE 3.

GRAF depletion attenuates myoblast differentiation by increasing Rho/ROCK activity. A, C2C12 cells were transfected with control or a mixture of two GRAF1-specific siRNAs, exposed to LSM for the indicated times, and cell extracts were immunoblotted with indicated antibody. Tubulin is shown as a loading control. Data are representative of three separate experiments. B, C2C12 cells were transfected with control or GRAF1-specific siRNAs and exposed to LSM for 72 h, and RhoA activity was measured an ELISA-based RhoA activity assay. Data represent the mean ± S.D. for three separate experiments. C, C2C12 cells were transfected with control or a mixture of two GRAF1-specific siRNAs, exposed to LSM with or without the ROCK inhibitor, Y27632 (10 μm), for the indicated times, and cell extracts were immunoblotted with indicated antibody. Tubulin is shown as a loading control. Data are representative of three separate experiments.