Fig. 6: Effect of global NQO1 overexpression and diet composition on the lysine acetylome in skeletal muscle and on enzymatic activities impacting acetyl-lysine turnover in liver extracts.

a Volcano plots of the lysine acetylome in skeletal muscle performed in the WT and NQO1-Tg mice fed either SD or HFD for 16 weeks. Graphs show the log2 fold-change of lysine acetylation between the indicated experimental groups of mice. n = 3 mice per group. b Venn-diagrams depicting the distribution of lysine-acetylated proteins in skeletal muscle of WT and NQO1-Tg mice on SD or HFD diet in response to the “TG-WT” and “HFD-SD” pairwise comparisons. Shared elements constitute attributes that are either diet- or genotype-independent, as indicated. Note that the digestion of a few proteins (e.g., myosin 1 and myosin 4) generated several peptides with the same acetylation site without and with other modifications such as peptide deamination and methylation. For this reason, these “shared” peptides were counted as one. Red font, hyperacetylation; blue font, hypoacetylation; black font, acetylation going in opposite direction. c Top pathways calculated by Ingenuity Pathway Analysis (IPA) using enrichment p-values. Bias-corrected z-score [positive (purple box), negative (green box), non-significant (white box)] are depicted as well as pathways without available pattern (gray box). d Normalized intensity values for a select group of acetyl peptides differentially influenced by diet x genotype interaction are depicted as boxplots (n = 3 per group). The lysine modification sites are shown. These data were analyzed using a two-way ANOVA. See Table S4 for complete list of acetyl peptides. e Heatmaps visualization of a select group of acetylated proteins in skeletal muscle that are known targets of the SIRT3 deacetylase. Left panel, effect of NQO1 overexpression; right panel, effect of diet. Upregulation (red), downregulation (blue). f Visualization of six proteins containing multiple lysine acetylation sites whose profiles are altered by genotype or diet in the indicated pairwise comparisons. g SIRT3, GOT2 and CRAT activities were performed in liver extracts. The data were analyzed using a two-way ANOVA. *p < 0.05; **p < 0.01.