Fig. 4. Impaired energy expenditure, and decreased lipolysis and mitochondrial activity in BAT from Ncc^−/− and Il18r^−/−Ncc^−/− mice in response to CL316243.

a–c Mouse metabolic parameters, including oxygen consumption (VO₂) (a), carbon dioxide production (VCO₂) (b), and energy expenditure (c) and their average values from full day cycle, light cycle, and dark cycle during 48 hrs of monitoring in WT, Ncc^−/−, Il18r^−/−, and Il18r^−/−Ncc^−/− mice after 7 days of CL316243 treatment (WT: n = 13; Ncc^−/−: n = 5, Il18r^−/−: n = 6; Il18r^−/−Ncc^−/−: n = 8). d. Representative images of UCP1 immunostaining of BAT sections and UCP1-positive area in EAT, SAT, and BAT from indicated groups of mice (WT: n = 7; Ncc^−/−: n = 5, Il18r^−/−: n = 5; Il18r^−/−Ncc^−/−: n = 6). Scale: 50 μm, inset: 25 µm. e Immunoblot analysis and quantification of UCP1 and PGC1α relative to GAPDH in BAT from indicated groups of mice (n = 3). f RT-PCR analysis of thermogenic, mitochondrial genes, lipogenic genes, lipolytic genes, M2 macrophage markers, eosinophil markers, and type 2 cytokines in BAT from indicated groups of mice (WT: n = 8; Ncc^−/−: n = 5, Il18r^−/−: n = 6; Il18r^−/−Ncc^−/−: n = 8). g RT-PCR analysis of BAT mitochondrial DNA (mtDNA) contents (mtDNA/nuclear (n)DNA ratio) in indicated groups of mice (WT: n = 5; Ncc^−/−: n = 4, Il18r^−/−: n = 3; Il18r^−/−Ncc^−/−: n = 3). h Immunoblot analysis and quantification of pHSL and Cyt C relative to total HSL or GAPDH in BAT from indicated groups of mice (n = 3). Data are mean ± SEM, one-way ANOVA test, followed by LSD post-test. Sample sizes were all biologically independent samples.