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. 2010 Oct 13;60(1):258–268. doi: 10.2337/db10-0281

FIG. 3.

FIG. 3.

Adenoviral overexpression of FTO in differentiated myotubes. Human myotubes or C2C12 cells were infected with recombinant adenovirus encoding human FTO or GFP (control) for 48 h. A: Representative Western blots of FTO, pSer473PKB, PKB, pPDK1, PDK1, and tubulin, in GFP- or FTO-overexpressing C2C12 myotubes. B: Representative Western blots of pSer473PKB and PKB total, in GFP- or FTO-overexpressing C2C12 myotubes (210 [7] ifu/well), under basal conditions or after insulin stimulation. Histogram represents means ± SEM (n = 4). *P < 0.05 versus basal situation, #P < 0.05 FTO versus GFP. C: Representative Western blots of FTO, pSer473PKB, PKB, p70/85S6K, and actin in human myotubes overexpressing either GFP or FTO (210 [7] ifu/well). D and E: Myotubes were transfected with siRNA control or specific for FTO for 48 h. D: Validation of FTO silencing in muscle cells. E: Representative Western blots of pSer473PKB and PKB in myotubes silencing for FTO (50 nmol/l of siRNA). Histogram illustrates the quantification and normalization of the phosphorylation of PKB in myotubes silenced for FTO. Values are means ± SEM (n = 3). ifu, inclusion-forming units.