Figure 4.

Effects of Ca²⁺ on activation of ERK1/2 and mTORC1. A, Cells were preincubated in KRBH with 4.5 mM Glc for 2 hours, treated with 100 μM diazoxide (DZ) or vehicle (DMSO) for 15 minutes, and then stimulated with 100 μM carbachol (Carb), AAs, or 100 nM EXD-4 for 2 minutes prior to harvest. One of four experiments is shown. B, Cells treated as in panel A were stimulated with AAs, 100 nM GLP-1, or 100 ng/mL EGF as indicated for either 2 or 30 minutes. One of two experiments is shown. Numbers under blots are fold change as in Figure 1. C, Cells were placed in fresh KRBH with or without Ca²⁺ plus 2 mM EGTA for 5 minutes before stimulation with AAs for 30 minutes. One of five experiments is shown. D, Cells preincubated in KRBH were stimulated with AAs or 100 μM Carb for the indicated times. pS6K and pS6 (S235/236) were immunoblotted. One of two experiments is shown.