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. 2016 Mar 1;30(4):402–416. doi: 10.1210/me.2015-1275

Figure 6. SIRT1 inhibition by SNP enhances insulin-induced Foxo1 phosphorylation (pFoxo1) and POMC expression.

Figure 6.

A and B, Pretreatment of mHypoA-POMC/GFP-2 cells with SNP (100μM, 1 h) before the addition of insulin (10nM, 15 min) significantly enhanced AKT phosphorylation (pAKT) and pFoxo1 as shown by Western blotting. C, Cotreatment of mHypoA-POMC/GFP-2 cells with insulin (10nM) and SNP (100μM) for 4 hours significantly up-regulated POMC mRNA relative to either treatment alone. D and E, Enhanced insulin-dependent changes in POMC mRNA in the presence of SNP were not due to changes in expression of the insulin receptor (Ins Rec) as shown by Western blotting (H2O: 1.79 ± 0.10 Ins Rec/β-actin; SNP: 1.88 ± 0.16; n = 3; P = .66). F, ChIP analysis of Foxo1 occupation of the POMC promoter indicating reduced and enhanced Foxo1 binding at 1 and 4 hours of SNP exposure, respectively (H2O: 0.009 ± 0.001; SNP 1 h: 0.003 ± 0.0006; SNP 4 h: 0.022 ± 0.008 signal relative to input; n = 3–11). H3+, histone 3a positive control.