FIG. 3.

Phosphomimetic mutant AKT(S473D) maintains C2C12 differentiation when rictor is downregulated. (A) C2C12 cells were transiently transfected with a vector control or one of 11 plasmids encoding constitutively active AKT(S473D), respectively, for 24 h, and protein expression was determined by immunoblotting with anti-Myc tag antibody. (B) Cells were then infected with lentiviruses encoding rictor shRNA or control shRNA for 24 h and then incubated in DM for 1 to 4 days. Representative microscopic fields from days 2 and 4 are presented. (C) MyHC was detected by immunofluorescence staining. (D) Quantitation of MyHC-positive (black bars) and multinucleated (open bars) cells was performed as described for Fig. 2F.