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. 2019 Jan 25;33(4):5654–5666. doi: 10.1096/fj.201802145RR

Figure 5.

Figure 5

Muscle-specific TRB3 overexpression does not augment the effect of food deprivation on protein synthesis rate and signaling pathway. TA muscles from groups with access to food (Fed) and food-deprived (FD) TG and WT littermate control mice were processed for analysis. A) Western blot with antipuromycin antibody was used to analyze puromycin incorporation into nascent peptides in WT and TG mice. Ponceau S staining was used to demonstrate an equal loading of protein. Methodology details can be found in Materials and Methods. B) The amount of puromycin incorporation was normalized by ponceau S staining. C–F) The phosphorylation and total expression of mTOR (C), 4EBP1 (D), PRAS40 (E), and TSC2 (F) were measured by Western blot analysis. A.U., arbitrary unit; p4EBP1S65, eukaryotic translation initiation factor 4E-binding protein 1; pmTORS2448, mechanistic target of rapamycin; PRAS40T246, proline-rich Akt substrate of 40 kDa; pTSC2T1462, tuberous sclerosis complex 2. Data are the means ± sem (n = 6–9/group). *P < 0.05, ***P < 0.001 vs. Fed. ##P < 0.01 vs. WT, ###P < 0.001 vs. WT.