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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Cell Metab. 2013 Oct 1;18(4):10.1016/j.cmet.2013.09.004. doi: 10.1016/j.cmet.2013.09.004

Figure 6. Serum from resveratrol-treated monkeys on HFS diet improves insulin signaling in 3T3-L1 adipocytes.

Figure 6

(A) IRS-1 protein levels in 3T3-L1 adipocytes incubated for 24-h with media containing serum from SD and HFS +/− Resv diet-fed monkeys for 2 years (n=3). Lanes were run on the same gel but were noncontiguous. (B) Phosphorylated Akt/Akt ratio in 3T3-L1 adipocytes pretreated for 24-h with media containing serum from SD and HFS +/− Resv diet-fed monkeys for 2 years and stimulated with insulin (100 nM) for 10, 20 and 30 min (n=3). (C) Number of cells expressing GLUT4 at the plasma membrane. 3T3-L1 adipocytes were incubated for 24-h with media containing serum from SD and HFS +/− Resv diet-fed monkeys for 2 years and then treated in the absence (0 min) or presence of 100 nM insulin (30 min). GLUT4-labeled sections of 3T3-L1 adipocytes are shown. Images were captured at 10× magnification. Scale bar: 90 μm. The number of cells that were stained for GLUT4 at the plasma membrane over total cell number both in control and insulin-treated cells were counted (n=5). (D) Insulin-induced cell surface GLUT4 labeling. 3T3-L1 adipocytes were pretreated for 24-h with media containing serum from SD and HFS +/− Resv diet-fed monkeys for 2 years and then stimulated with insulin (100 nM) for 30 min. GLUT4-labeled sections of 3T3-L1 adipocytes are shown. Images were captured at 60× magnification. Scale bar: 15 μm. The amount of GLUT4 labeling at the plasma membrane of insulin-treated cells was normalized by the total GLUT4 staining in the same cells (n=5). (A to D) The graphs show the mean ± SEM, and the results are expressed as percent increase over the values observed in adipocytes treated with HFS serum (A and D) or as percent increase over the values observed in adipocytes at baseline (B and C). The data were analyzed using One-Way ANOVA and RM-ANOVA was used to calculate the time effect (P time), the diet effect (P diet) and the diet × time interaction (P diet × time). *, P < 0.05 (HFS vs. HFS + Resv serum); #, P < 0.05 (HFS vs. SD serum). SD: standard diet; HFS: high-fat, high-sugar; Resv: resveratrol; pAkt: phosphorylated Akt; PM: plasma membrane.