Myf5 translocation to a cytoplasmic
compartment in RhoG-, Rac1-, and Cdc42Hs-expressing cells and after JNK
activation. (A) Protein extracts from parental L6, L6 RhoGV12, L6
Rac1V12, and L6 Cdc42HsV12 (100 μg/well) were
immunoblotted with anti-Myf5 antibody as described in
MATERIALS AND METHODS. (B) Parental L6 (a and b), L6 RhoAV14 (c), L6
RhoGV12 (d), L6 Rac1V12 (e), and L6 Cdc42HsV12 (f) were fixed and
stained for Myf5 expression. Panels a, c, d, e, and f, cells in
proliferating medium; panel b, 1.5 d after the addition of DM. For
each panel, cells shown are representative of five independent
experiments with more than 100 observed cells. Bar, 10 μm. (C)
Parental L6 (a) and L6 RhoAV14 (b) were treated with anisomycin (50 nM)
for 24 h and then fixed and stained for Myf5 localization. L6
myoblasts were transfected with constructs expressing FLAG-tagged MKK7
or HA-tagged MKK3. Twenty-four hours after transfection, cells were
fixed and processed for FLAG (c, inset) or HA epitope detection (d,
inset) and for Myf5 localization (c and d). For each panel, cells are
representative of three independent experiments.