Fig. 7
Effects of insulin on ERK and Akt phosphorylation in GFP + Shb-RIN cells. Sorted or nonsorted GFP-RIN or GFP + Shb-RIN cells (2 × 105) were serum deprived 2 days after transfection for 60 min followed by stimulation with 100 ng/ml insulin for 10 min. Cells were lysed in SDS-β-mercapthoethanol buffer, run on 9% SDS-PAGE gels, and immunoblotted for phospho-ERK, total ERK, phospho-Akt, and total Akt expression. Filters were stripped in SDS-β-mercaptoethanol solution between blotting with the two different antibodies. Results from three separate experiments are expressed as means ± SEM. *p < 0.05 using Students paired t-test.