Figure 3
MyoD-/-myoblasts maintain cyclin D expression after serum withdrawal. (A, C) RNA was isolated from a differentiation time course of WT and MyoD-/- primary myoblasts and an RNase protection assay was performed with the indicated probes. The protected 32P-labeled fragments were separated on a polyacrylamide gel, the gel dried and exposed to X-ray film. (B, D) The amount of each protected fragment was quantitated using a phosphorimager. The values obtained were normalized relative to the GAPDH control and the L32 control, and graphed as a function of the number of days in differentiation medium (DM), n = 3. (E) Nuclear protein was isolated from a differentiation time course of WT and MyoD-/- primary myoblasts and a western blot was performed with the indicated antibodies. (F) Quantitative real-time PCR for Cyclin D3 of WT and MyoD-/- myoblasts, n = 3.