Effect of RhoGV12, Rac1V12, Cdc42HsV12, and
RhoAV14 on muscle-specific protein expression. (A) Proliferating
parental L6 and L6 RhoGV12, L6 Rac1V12, L6 Cdc42HsV12, and L6 RhoAV14
myoblasts were grown to ∼80% and shifted to DM for 1 d. Cells
were then fixed and stained for myogenin expression and nuclei with
Hoechst. The histogram represents the percentage of myogenin-positive
cells and summarizes the data from five independent sets of
experiments; 40–50 cells were analyzed in each experiment. (B) Protein
extracts from parental L6, L6 RhoGV12, L6 Rac1V12, L6 Cdc42HsV12, and
L6 RhoAV14 (100 μg/well) collected at the indicated periods (P,
proliferative; D1, D2, D3, D4, 1–4 d after addition of differentiation medium) were
immunoblotted with an anti-troponin T antibody as described
in MATERIALS AND METHODS. Shown are Western blots indicating the
expression of troponin T in parental L6 (a), L6 RhoGV12 (b), L6 Rac1V12
(c), L6 Cdc42HsV12 (d), and L6 RhoAV14 (e). (C) Proliferating parental
L6 (a) and L6 RhoGV12 (b), L6 Rac1V12 (c), and L6 Cdc42HsV12 (d)
myoblasts were grown to ∼80% and shifted to DM for 3 d. Cells
were then fixed and stained for MHC expression. For each panel, cells
shown are representative of five independent experiments with more than
100 observed cells. Bar, 10 μm.