Extended Data Fig. 1: Loss of endogenous ketogenesis in Hmgcs2-deficient mice impairs cold-induced beige fat formation and mitochondrial biogenesis.
a. Hepatic Hmgcs2 mRNA in control (CTL) and Hmgcs2 knockout (KO, Hmgcs2−/−) male mice at P0, P7, P14, and P21. b. Serum βHB levels in CTL and KO mice at P0, P4, P7, P14, and P21. c. Survival rate of Hmgcs2 heterozygous (HET) and KO mice (n = 42, 16). d. Ucp1 mRNA in iWAT of wild-type mice after 1, 2, 5, or 7 days at 4 °C (n = 3/group). e. Hmgcs2 mRNA in iWAT of wild-type mice after 7 days at 4 °C (n = 5/group). f. iWAT weights of CTL and KO mice housed at room temperature (RT) or 4 °C (Cold) for 7 days. g. Quantification of white and beige adipocytes from iWAT sections in Fig. 2c (n = 3/group). h–j. Transcriptomic analysis of iWAT SVF: pie charts (top) and volcano plots (bottom) of differentially expressed genes for (h) CTL Cold vs. RT; (i) KO Cold vs. RT, and (j) KO vs. CTL under Cold conditions. k. Ucp1 and Cidea mRNA in iWAT of mice from (f). l. iBAT weights in mice from (f). m. Ucp1 mRNA in iBAT from mice in (f). n. Fatty acid oxidation (FAO) stress test in SVF-derived adipocytes from CTL and KO mice using palmitic acid (PA)-BSA as substrate, followed by etomoxir (Eto) treatment. Real-time oxygen consumption rate (OCR) shown (n = 3 for CTL; 4 for KO). o. Quantification of basal and maximal FAO responses following Eto treatment (n = 3 for CTL; 4 for KO). p. Mitochondrial DNA (mtDNA) copy number in iWAT, expressed as the ratio of Nd1 to Lpl gene expression (n = 6/group). q. mRNA levels of mitochondrial biogenesis genes in iWAT. Data are expressed as mean ± SD. n = 4/group in a–b, f, k–m, q. Statistical analyses: two-sided Student’s t-test (a, b, d, e, o–q) and two-way ANOVA with Tukey’s post hoc test for multiple comparisons (f, k–m) and. P values in (d) are versus day 0.
