Fig. 3: Acod1 loss improves metabolic activity of HFD-fed mice.

Whole-body O₂ consumption rate (VO₂) (A) and energy expenditure rate (B) normalized on body weight of HFD-fed WT (n =n = 6) and Acod1^-/- (n = 8) mice during light and dark hours (one representative experiment out of two independently executed). C Venn diagram (left) and gene ontology (biological processes)-based functional enrichment analysis (right) of downregulated proteins in eWAT of WT mice fed with HFD for 16 weeks compared with ND-fed mice (HFD vs ND [WT]) and rescued by Acod1 loss (Acod1^-/- vs WT [HFD]). Data derive from n = 4 (ND-fed WT) and n = 5 (HFD-fed) mice from one experiment. D Heatmap depicting mRNA levels of selected genes related to tricarboxylic acid (TCA) cycle, oxidative phosphorylation (OXPHOS), lipolysis and fatty acid (FA) oxidation downregulated in eWAT of HFD-fed WT mice (HFD vs ND [WT]) and rescued by Acod1 loss (Acod1^-/- vs WT [HFD]) Data derive from mice described in C: n = 4 (ND-fed WT) and n = 5 (HFD-fed) mice from one experiment. E Acyl-carnitine levels in iWAT of mice fed with either ND (n = 6) or HFD (n = 10) from two independent experiments. Carn.: carnitine (F) qPCR analysis of mRNA levels of the indicated genes in BAT of mice described in A; n = 10 per group from two independent experiments. G qPCR analysis of Ucp1 mRNA levels in BAT of WT and Acod1^-/- mice fed with either ND or HFD for 16 weeks. n = 7–9 per group from two independent experiments. H Representative images of UCP1 immunohistochemical staining normalized to cell number in BAT of mice described in G. Bar = 100 μm. I qPCR analysis of Ucp1 mRNA levels in iWAT of mice described in A; WT (n = 15), Acod1^-/- (n = 14) mice from three independent experiments. J Heatmap depicting mRNA levels of selected genes known as inducers (positive regulators) or inhibitors (negative regulators) of adaptive thermogenesis changed in iWAT of HFD-fed WT mice and rescued by Acod1 loss. Data derive from n = 3 (ND-fed WT), n = 5 (HFD-fed WT), and n = 3 (HFD-fed Acod1^-/-) mice from one experiment. In: (B), (E, F), (G, I) data as mean±s.e.m; (A) two-way ANOVA; (E, G) one-way ANOVA followed by Dunnett’s multiple comparisons test compared to HFD-fed WT mice; (B, F, I), t test; *P < 0.05; **P < 0.01; ***P < 0.001.