Figure 6

Ca _V 1.1/CaMKII modulates Ras/Raf/ERK1/2 signaling to increase mTORC1 signaling in response to electrical stimulation. (A) Representative western blot for activated Ras (Ras-GTP) in Raf-RBD pull-down from WT and EK muscle. (B) Analysis of activated Ras (Ras-GTP) in Raf-RBD pull-down in muscle from WT and EK mice. (C) Electrical stimulation-induced changes in mTORC1 signaling. Representative western blots of isolated solei and EDL muscles subjected to a five-minute electrical stimulation (as described in Methods) in the presence and absence of KN-93 (5 μM). For all analyses the values for the western blots of individual bands from WT and EK muscle are each normalized to the values obtained in the absence of electrical stimulation. For statistical analyses the WT and EK are each compared to their unstimulated controls. (D) Analysis of pCaMKII/GAPDH in the soleus. (E) Analysis of pCaMKII/GAPDH in the EDL. (F) Analysis of p-Raf (a CaMKII target) in the EDL. (G) Analysis of pERK1/2/GAPDH in the soleus. (H) Analysis of pERK1/2/GAPDH in the EDL. (I) Analysis of pS6/S6 in the soleus. (J) Analysis of pS6/S6 in the EDL. (K) Analysis of p4EBP1/4EBP1 in the soleus. (L) Analysis of p4EBP1/4EBP1 in the EDL. (M) Summary of changes in signaling pathways. Values are shown as mean ± SEM. *P <0.05, **P <0.01, and ***P <0.001.