Figure 3
JNK inhibition by C66 attenuated insulin resistance induced by prednisone or 11β-HSD1 overexpression in adipocytes.
(a) 3T3-L1 cells differentiated into adipocytes and then treated with PF00915275 (10 μM), C66 (10 μM) or vehicle (DMSO) for 2 h followed by incubation with prednisone (5 μM) or DMSO for 6 h. Glucose uptake rate was measured as described in the methods section. (b) Cells were treated with PF00915275 (10 μM), C66 (10 μM) or vehicle (DMSO) for 2 h and then with prednisone (5 μM) or DMSO for 2 h. p-Akt/Akt proteins levels were then determined by western blotting. The gels were run under the same experimental conditions. Shown are cropped gels/blots (The gels/blots with indicated cropping lines are shown in Supplementary Fig. S1). (c–f) Cells were treated as in A and mRNA levels of IRS-1 (c), GLUT4 (d), Adiponectin (e) and FABP (f) were examined by qPCR. (g) Cells were treated as in B and protein levels of p-IRS1Ser302 and IRS1 were examined by western blot analysis. The gels were run under the same experimental conditions. Shown are cropped gels/blots (The gels/blots with indicated cropping lines are shown in Supplementary Fig. S1). Representative blots from 3 independent experiments were shown. (h,i): Differentiated 3T3-L1 cells with or without 11β-HSD1 overexpression were treated with C66 (10 μM) or vehicle (DMSO) for 2 h and then incubated with insulin (100 nM) for 15 min. Glucose uptake rate (h) and Akt phosphorylation (i) were determined. The gels were run under the same experimental conditions. Shown are cropped gels/blots (The gels/blots with indicated cropping lines are shown in Supplementary Fig. S1). *P < 0.05, **P < 0.01, ***P < 0.001; all data from 3 independent experiments.
