GAS muscles were collected from WT and TRB3TG mice. (A–D) The activations of signaling molecules related to protein synthesis, including mTOR (A), S6K1 (B), and Akt (C and D), were analyzed by Western blot. (E–G) After 5 hours of fasting, mice received an intraperitoneal injection of 0.04 µmol/g body weight with puromycin dissolved in 100 µl of PBS 30 min before euthanasia to measure the rate of protein synthesis. Western blot was used to measure the amount of puromycin incorporation into nascent peptides. (E) Ponceau S staining of puromycin analysis from SOL muscle showed that proteins were equally loaded. EDL (F) and SOL (G) muscles were analyzed. Data are the means ± S.E.M. (n = 5–6 per group). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. WT