Fig. 6: Hepatic Mc3r reactivation restores adaptative energy expenditure, cellular respiration, and lipid droplet autophagy.

a Energy expenditure (kcal/mouse/h) at 6 °C from Mc3r^+/+(n = 15), Mc3r^TB/TB (n = 16), and Mc3r^Hep/Hep (n = 12) independent observations. b EE (kcal/mouse/h) at 6 °C from Mc3r^+/+ (n = 15), Mc3r^TB/TB (n = 16), and Mc3r^Hep/Hep (n = 12) independent observations. c Oxygen consumption (VO₂) (ml/mouse/h) at 6 °C from Mc3r^+/+ (n = 15), Mc3r^TB/TB (n = 16), and Mc3r^Hep/Hep (n = 12) independent observations. d VO₂ (ml/mouse/h) at 6 °C from Mc3r^+/+ (n = 15), Mc3r^TB/TB (n = 16), and Mc3r^Hep/Hep (n = 12) independent observations. e Oxygen consumption rate (OCR) from cultured primary hepatocytes from Mc3r^+/+, Mc3r^TB/TB and Mc3r^Hep/Hep from 3 independent experiments. f Basal respiration OCR (pmol/min/5000 cells) from 3 independent experiments. g Maximal respiration OCR (pmol/min/5000 cells) from 3 independent experiments. h Bulk RNA-Sequencing analysis of mouse liver samples from Mc3r^+/+, Mc3r^TB/TB, and Mc3r^Hep/Hep genotypes, all on a normal chow diet (n = 4) independent observations. Pairwise scatter plot illustrates the Log₂ fold change (Log₂FC) values for all significantly differentially expressed genes across three genotype comparisons. The Log₂FC for Mc3r^TB/TB vs Mc3r^+/+ is plotted against Mc3r^Hep/Hep vs Mc3r^+/+ (blue-filled circles), and linear regression fit lines are shown for the comparisons between Mc3r^TB/TB vs. Mc3r^+/+ and Mc3r^Hep/Hep vs Mc3r^+/+ (blue line). i The Log₂FC for Mc3r^TB/TB vs Mc3r^+/+ is plotted against Mc3r^Hep/Hep vs Mc3r^TB/TB (orange-filled circles). Linear regression fit lines are shown for the comparisons between Mc3r^TB/TB vs. Mc3r^+/+ and Mc3r^Hep/Hep vs Mc3r^TB/TB (orange line), with the black line shows best fit perfect correlation lines included for visualization. j Heatmap depicting significant gene ontology (GO) biological pathways related to lipid droplet autophagy, lipid metabolism, and energy homeostasis. The negative log₁₀ P values for these pathways were compared across the Mc3r^TB/TB vs. Mc3r^+/+, Mc3r^Hep/Hep vs Mc3r^+/+ and Mc3r^Hep/Hep vs Mc3r^TB/TB comparisons (adjusted p-value ≤ 0.05). k Western blot images of liver lysates from Mc3r^+/+, Mc3r^TB/TB and Mc3r^Hep/Hep. l Quantification of LC3II normalized by β-actin in (g) from Mc3r^+/+ (n = 12), Mc3r^TB/TB (n = 10), and Mc3r^Hep/Hep (n = 11) independent observations. m Quantification of LC3II normalized by LC3I in (g) from Mc3r^+/+(n = 12), Mc3r^TB/TB (n = 10), and Mc3r^Hep/Hep (n = 11) independent observations. Data are represented as mean ± SEM. Groups were compared by one-way ANOVA followed by Fisher’s LSD test (b, d), one-way ANOVA followed by Tukey’s HSD test (f, g), two-tailed simple linear regression (h, i). p < 0.05; ** p < 0.01; *** p < 0.001. Source data are provided as a Source Data file.