Figure 2.

LY294002 inhibits MyoD-mediated differentiation. Results of time-course experiments are shown for C2BP5 cells infected with Ad:MyoD and incubated in DM with vehicle (dimethyl sulfoxide, DMSO) or the PI3-kinase inhibitor LY294002 in DMSO (LY, 20 μM) for 1 or 2 d. (A) Results of immunocytochemistry for MHC (red) and myogenin (green). Magnification, 200×. (B) Top, quantification of myotube area by using MHC staining (mean ± SEM of three experiments counting 20 fields at 200× magnification). Bottom, quantification of myogenin expression (mean ± SEM of 12 fields from two experiments at 200× magnification). (C) Immunoblots for MyoD, myogenin, MHC, troponin T, Akt, phospho-Akt (Ser 473) (p-Akt), and Akt substrates (Akt sub) by using whole cell protein lysates.