Fig. 2: Nitro-oleic acid treatment alters myocardial proteome and glucose metabolism in HFpEF mice.
a Principal component analysis (Chow: N = 9; HFD + L-NAME vehicle: N = 8; HFD + L-NAME NO2-OA: N = 6). Volcano-plots comparing high-fat diet (HFD) + L-NAME vehicle with chow mice (b) and NO2-OA-treated with vehicle-treated HFD + L-NAME mice (d). Significantly changed proteins are marked (significance ≥ -log10(0.05); red: fold change > log2(1.5); blue: fold change < -log2(1.5)). Protein names, log fold changes and p-values are listed in the source data file. c KEGG pathway enrichment analysis of significantly increased proteins of HFD + L-NAME vehicle compared to chow mice (protein count ≥ 2; significance ≥ -log10(0.05)). e Venn diagram of proteins significantly decreased in the HFD + L-NAME vehicle compared to the chow group and proteins significantly increased in NO2-OA-treated compared to vehicle-treated HFD + L-NAME mice. f Body weight (N = 12/12/12) and glucose tolerance test (N = 7/7/7). g Protein abundances of solute carrier family 2, member 4 (GLUT4) and pyruvate dehydrogenase kinase 4 (PDK4) relative to cytochrome c oxidase 4 (COX4) derived from liquid chromatography–mass spectrometry (N = 8/8/6). h Quantification of immunoblots (IB) for PDK4 relative to COX4 and phosphorylated pyruvate dehydrogenase (p-PDH) relative to COX4 (N = 9/8/6). Protein levels were normalized to the total protein amount assessed on IB by fluorescence labeling (N = 9/8/6) (Fig. S12 and S13). Data are presented as mean ± standard deviation for f (left), g and h. f (right) shows mean ± standard error of the mean. Data in g and h are shown relative to the mean of the chow group. Statistical significance for f (left), g and h (left) was calculated by One-way ANOVA followed by Bonferroni´s post-hoc test. Kruskal-Wallis-test followed by Dunn´s multiple comparison test was used for h (right). Two-way ANOVA followed by Bonferroni´s multiple comparisons test was used for f (right). * indicates differences between HFD + L-NAME vehicle and HFD + L-NAME NO2-OA. Statistical significance was calculated with Perseus (MPI Biochemistry) for b and d and with two-tailed Fisher’s exact test for c. Only statistically significant differences are indicated. p *<0.05, **<0.01, ****<0.0001. Outliers in g were identified using ROUT method and excluded from analysis.
