Fig. 4
From: Sirt2 facilitates hepatic glucose uptake by deacetylating glucokinase regulatory protein
Sirt2 deacetylates GKRP. a–d Immunoblot analysis (IB) with the indicated antibodies of total cell lysate and precipitates after immunoprecipitation (IP) with anti-acetylated lysine (AcK) antibodies from HEK293 cells expressing Flag-tagged GKRP and Flag-tagged Sirt2 with and without nicotinamide (NAM), AGK2, Ex527, or trichostatin A (TSA) (a), from Sirt2 knockdown primary hepatocytes cultured in low- or high-glucose medium (b), from db/db mouse-derived hepatocytes with and without adenovirus-mediated overexpression of Sirt2 cultured in low- or high-glucose medium (c), and from db/db mouse-derived hepatocytes with NMN treatment and with and without Sirt2 knockdown cultured in high-glucose medium (d). The quantification of acetylated GKRP levels is normalized to GKRP expression in the right graph (b–d, n = 3 each). e, f Acetylated GKRP levels in the liver of mice fed normal chow (NC) or a high-fat diet (HFD) with adenovirus-mediated hepatic overexpression of Sirt2 after 16-h fasting (e) and acetylated GKRP and keratin 8 levels in the liver of Sirt2 knockdown lean mice after 16-h fasting (f). g, h Successive comparison of lysine acetylation levels of keratin 8 and GKRP in the liver of NC- or HFD-fed mice after NMN injection after 16-h fasting (g) and in the liver of NC-fed mice treated with siSirt2 at 30 min after administration of glucose (4 g/kg) after 16-h fasting (h). *P < 0.05; one-way ANOVA with the Fisher’s PLSD post-hoc test (b–d). All data are representative of at least three independent experiments. Error bars show s.e.m
