Changes in mTORC1-dependent signaling in denervated muscles. (A, D) Western blot analysis of tibialis anterior (TA) and soleus (Sol) muscles after six days of denervation using antibodies against PKB/Akt phosphorylated at Serine 473 (A) and S6 phosphorylated at Serines 235/236 (D). α-actinin was used as a loading control. (B, C, E, F) Relative mRNA levels of atrogin-1/MAFbx (Atr-1) and MuRF1 as determined by qPCR in TA and soleus (Sol) muscles after six days of denervation. Note that expression of both E3 ligases is blunted in RAmKO mice (B, C), while this response is exaggerated in TA (E) but not in soleus muscles (F) of TSCmKO mice. (G – J) Relative mRNA levels of Pgc1α and Pgc1β in RAmKO (G, H) and TSCmKO (I, J) mice after six days of denervation. All values are normalized to the expression levels of the transcript measured in innervated muscle of control littermates (set to 100%). N ≥4 mice for TA and N ≥5 mice for soleus of each genotype. Values represent mean ± SEM. P-values are ***P <0.001; **P <0.01; *P <0.05; Student’s t-test.