Fig 2: Feeding-induced activation of mTORC1 and translation of theRps8 mRNA are blunted in the livers of rats fed an HFD compared with a CON diet. Representative blots showing phosphorylation of p70S6K1 on Thr389 (A) and 4E-BP1 on Ser65 (B). (C) The proportion of ß-actin andRps8 mRNAs in polysomes. Values are means ± SEMs;n = 10 (polysome analysis) orn = 15–16 (p70S6K1 and 4E-BP1 phosphorylation). *Different from CON,P < 0.01. CON, control diet; HFD, high-fat diet; mTORC1, mechanistic (mammalian) target of rapamycin in complex 1; -P, phosphorylated; p70S6K1, 70-kDa ribosomal protein S6 kinase 1; Rps8, ribosomal protein S8; S65, Ser65; -T, total; T389, Thr389; 4E-BP1, eukaryotic initiation factor 4E binding protein 1.

Fig 3: Effect of cast immobilization on phosphorylation of p70S6K1 and 4E-BP1. Phosphorylation of (A) p70S6K1 on Thr389 and (B) 4E-BP1 on Ser65 was assessed by Western blot analysis of muscle from mice that had been immobilized, fasted overnight, and then re-fed for 30 min prior to sacrifice. The results are expressed as a ratio of the signal obtained for the phosphorylated protein relative to that for the total protein. Data displayed as cast immobilized (open bars) versus contralateral non-immobilized control (closed bars). The data are presented as means ± SE of the arbitrary density values (ADV); n = 6–7 mice/group (1 Fasted/Non-Immobilized outlier removed). Groups sharing a similar letter are not significantly different (P = 0.05).