FIG 8.

Overexpression of β-catenin restores reduced cMYC and protein synthesis. (A) Representative immunoblotting and densitometry of total (T) and active (A) β-catenin, cMYC, and UBF1 in untreated (UnT) or 10 mM AmAc-treated murine C2C12 myotubes overexpressing wild-type β-catenin or a mock construct. (B) Representative immunoblotting and densitometry of ribosomal proteins RPL5, RPL23, and RPL32 in untreated C2C12 myotubes or those treated with 10 mM AmAc for 24 h, stably overexpressing either wild-type β-catenin or a mock construct. (C) Representative immunoblotting and densitometry of puromycin incorporation in untreated C2C12 myotubes or those treated with 10 mM AmAc for 24 h, overexpressing wild-type β-catenin or a mock construct. (D) Relative fold changes in 45S rRNA expression in untreated myotubes or those treated with 10 mM AmAc for 24 h, overexpressing wild-type β-catenin or a mock vector. (E) Representative photomicrographs and myotube diameters of differentiated C2C12 myotubes with β-catenin overexpression compared with mock vector-transfected myotubes treated with and without 10 mM AmAc. Scale bars, 40 μm. All cellular experiments were performed in at least 3 biological replicates, all blots were normalized to β-actin, and data are expressed as means ± SD. ***, P < 0.001 (mock versus β-catenin overexpression). The antibody against active β-catenin recognizes the unphosphorylated, stable form of β-catenin.