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Immunoblot analysis of GS and PP1 protein content. Cultured human myotubes were transduced with Ad-GFP, Ad-R6, Ad-PTG or Ad-GM and incubated with 25 mM glucose for 48 h. Immunoblot analyses were performed on cell extracts (20 μg protein for GS and phospho-GS and 5 μg protein for PP1). Membranes were hybridized with antibodies against: (A) phospho-GS (Ser-641/0), (B) muscle GS and (D) PP1α. (A, B, D) A representative image is shown. Bands were quantified with a LAS-3000 (FujiFilm). Data are means ± SEM from three experiments performed in duplicate. Significance of differences versus cells treated with Ad-GFP under the same incubation conditions: *p < 0.05 and **p < 0.01.