Fig. 4: Activation of the AMPK signaling pathway is induced by nitro-oleic acid in LV of HFpEF mice.
a KEGG pathway enrichment analysis of proteins derived from liquid chromatography followed by mass spectrometry (LC-MS) with significantly enhanced abundance after administration of NO2-OA in high-fat diet (HFD) + L-NAME treated mouse hearts compared to vehicle-treated mouse hearts (HFD + L-NAME vehicle) (inclusion criteria: protein count ≥ 10; pathway enrichment factor ≥ 30%; significance ≥ -log10(0.05)). The most significant KEGG pathway is marked with a red arrow. b Heatmap of all proteins associated with the AMP-activated protein kinase (AMPK) signaling pathway and identified by LC-MS analysis. Protein names and z-scores are in the source data file. c Representative immunoblots (IB) and quantification of AMPK and phosphorylation level of AMPK at threonine residue 172 (p-AMPK), sirtuin 1 (SIRT1) and sirtuin 3 (SIRT3) (N = 9/8/6). Protein levels were normalized to the total protein amount assessed on IB by fluorescence labeling (Figs. S12 and S14). Data in c are shown relative to the mean of the chow group as mean ± standard deviation. Statistical significance was calculated by two-tailed Fisher’s exact test for a and One-way ANOVA followed by Bonferroni´s post-hoc test for c. Only statistically significant differences are indicated. p: **<0.01, ***<0.001, ****<0.0001.
