Effect of rapamycin on mTORC1 and ATF4 after exercise and leucine. (A) Experimental set-up. (B–E) Bar graphs showing downstream kinases of mTORC1 in SOL; (B) quantification of pS6K1Thr389, (C) Quantification of pRPS6Ser235/236, (D) quantification of mTORSer2448, (E) quantification of 4E-BP1Ser65. (F) qPCR of ATF4- dependent genes upon exercise and acute rapamycin administration. (G) Quantification of LARS. (H) Quantification of ATF4. (I) Quantification of eIF2αSer51. (J) Representative immunoblots of downstream mTORC1 signaling and ATF4 pathway. Bars represent mean, circles represent individual values, all data is shown as fold change to Sed sal (dashed horizontal line). Panel B–E and G-I, (Sed sal n = 6), (Run0 n = 5), (Run0+rapa n = 5), (Run48 n = 5), (Run48+rapa n = 5). One-Way ANOVA with Tukey's multiple comparisons test. ∗ p < 0.05; ∗∗ p < 0.01, ∗∗∗p < 0.001. ∗∗∗∗p < 0.0001.