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. Author manuscript; available in PMC: 2016 Jan 31.
Published in final edited form as: Nat Med. 2015 Jul 13;21(8):887–894. doi: 10.1038/nm.3882

Figure 4. Nnmt regulates Sirt1 protein stability through its product MNAM.

Figure 4

(a) Sirt1 protein and mRNA expression in control and Nnmt overexpressing primary hepatocytes (representative of 3 independent experiments performed in triplicates). (b) Sirt1 protein and mRNA expression in control and Nnmt knockdown primary hepatocytes (representative of 3 independent experiments performed in triplicates). (c) Sirt1 protein and mRNA expression in livers of control and Nnmt knockdown mice (shcontrol n = 8, shNnmt n = 8). (d) Time course of Sirt1 protein degradation in cyclohexamide-treated control and Nnmt overexpressing hepatocytes (triplicate determination). (e) Correlation between Nnmt mRNA and Sirt1 protein expression from human liver biopsies (n = 12, P = 0.797). (f) Supplementation of media with 0.1 mM MNAM increases intracellular MNAM concentration (triplicate determination). (g) Dose-dependent effect of MNAM (0–1 mM) on Sirt1 protein expression in primary hepatocytes (representative of 2 independent experiments). (h) Dose-dependent effect of MNAM (0–0.3 mM) on G6pc ,Pck1, Sirt1 and Nnmt expression in primary hepatocytes (representative of 3 independent experiments performed in triplicates). (i) Glucose production in primary hepatocytes supplemented with MNAM and infected with either control or Sirt1 knockdown adenovirus (triplicate determination). (j) G6pc and Pck1 expression in primary hepatocytes supplemented with MNAM and infected with either control or Sirt1 knockdown adenovirus (triplicate determination). Data are presented as mean ± s.e.m. Statistical significance was tested by unpaired Student’s t-test. *P < 0.05. RU = relative units.