Extended Data Fig. 2: Global and liver-specific deletion of Hmgcs2 impairs cold-induced beige fat formation in male mice.
a. Hmgcs2 mRNA in liver, iWAT, eWAT, and iBAT of P14 control mice (n = 3/group). b. Intracellular βHB concentrations in liver, iWAT, eWAT, and iBAT of P14 control mice (n = 4/group). c. Expression of Hmgcs2 (left) and Fabp4 (right) during adipogenic differentiation of SVF (n = 3/group). d. Schematic of global and conditional Hmgcs2 knockout models and experimental design. Eight-week-old male control (CTL), global knockout (KO, mediated by Sox2-Cre), liver-specific knockout (LiKO, mediated by Albumin-Cre), and adipocyte-specific knockout (AdKO, mediated by Adipoq-CreERT2) mice were maintained on chow and housed at room temperature (RT) or 4 °C (Cold) for 7 days. Rectal temperature was measured after 2 days; iWAT was collected after 7 days. e. Representative western blot of HMGCS2 protein in liver and iWAT. β-actin as loading control. f. Serum βHB levels at P14 in male mice (n = 6 for CTL; 5 for KO; 4 for LiKO; 4 for AdKO). g. Intracellular βHB in iWAT at P14 (n = 3/group). h. Serum βHB levels at P14 in female mice (n = 8 for CTL; 7 for KO; 6 for LiKO; 6 for AdKO). i. Changes in rectal temperature after 2 days at 4 °C in male mice. Delta rectal temperature = temperature at indicated time – baseline temperature (n = 8 for CTL RT; 8 for CTL Cold; 6 for KO; 6 for LiKO; 5 for AdKO). j. Gross morphology and representative H&E-stained iWAT sections. Grid:1 cm×1 cm. Scale bar, 100 µm. k. Quantification of white and beige adipocyte in (j) (n = 4/group). l. mRNA levels of thermogenic genes in iWAT (n = 6 for CTL RT; 6 for CTL Cold; 4 for KO; 4 for LiKO; 4 for AdKO). Data are expressed as means ± SD. Statistical analyses: one-way ANOVA with Tukey’s post hoc test for multiple comparisons (a–b, f–i, l) and two-sided Student’s t-test (c). P values in (c) are versus day 0 (c). Illustrations in d created in BioRender.com, https://biorender.com/hjq5y60.
