Figure 4.

Western blot analysis of myogenin protein expression in C2C12 muscle cells during five subsequent days of experiment. From top to bottom: control, myxothiazol (1 µm), effect of chloramphenicol (0.1 mg/ml), insulin (1 nm), rotenone (10 µm) being present 30 min prior to insulin (1 nm) addition, or oligomycin (1 ng/ml) being present 30 min prior to insulin (1 nm) addition. Nuclear protein extracts (50 µg of protein per sample) isolated from treated (see the text) or not (Ctrl) mouse C2C12 muscle cells at each day of experiment were resolved by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) followed by Western blotting with rabbit antimyogenin polyclonal antiserum. After washing and additional blotting with secondary HRP‐conjugated donkey antirabbit antiserum, the blots were detected with ECL reagent (see Material and Methods). All antibodies were from Santa Cruz, CA. Each experiment was repeated at least twice with similar results.