Fig. 4: The sex-selective basal transcriptional activity of the beiging process in subcutaneous WAT is under Ncoa1 control.

a PCA of differentially expressed genes (DEGs) in SAT of WT females (n = 3) vs. WT males (n = 4) under basal (NaCl) conditions. b, c KEGG pathway enrichment bubble plots of DEGs in SAT of WT mice under basal conditions, comparing females (n = 3) vs. males (n = 4). Genes upregulated in WT females (b) and WT males (c). d RNAseq expression analysis of Elovl family genes in SAT of Ncoa1 KO and WT mice (n = 4 animals per group, except for females WT NaCl, Ncoa1 KO NaCl, and Ncoa1 KO CL: n = 3 per group). e Luciferase assays monitoring the transcriptional activity of the Ucp1 promoter (− 3750 / + 490 bp) in HeLa cells co-transfected with Ncoa1 and the indicated coregulators. Data are mean of 3 independent experiments, each performed in triplicates. f RNAseq expression analysis of Gata family genes in SAT of Ncoa1 KO and WT mice (same n as in (d)). g, h Luciferase assays monitoring the effect of Gata3 and Ncoa1 on Ucp1 promoter activity. Data are mean of 3 independent experiments, each performed in triplicates. i ChIP assay showing Ncoa1 and Gata3 binding to the proximal Ucp1 promoter in SAT of WT or Ncoa1 KO females and WT males. Data are mean of 3 independent experiments performed in 2 to 4 replicates. j GATA3 expression (RNAseq) in SAT and VAT from GTEx donors: women (SAT: n = 218; VAT: n = 170) and men (SAT: n = 445; VAT: n = 371). Data are in TPM (Transcript Per Million). k qRT-PCR analysis of Gata3 expression in SAT and BAT of female and male WT and Ncoa1 KO mice (n = 4 per group). l ATAC-seq at the Gata3 locus in both sexes (n = 2 WT mice per sex). Left: genome browser tracks; Right: average peak quantification. m–r Energy expenditure measured by indirect calorimetry during day (white) and night (gray) in WT and Ncoa1 KO mice: females (WT: n = 4; KO: n = 6) (m–o), and males (WT: n = 5; KO: n = 6) (p–r). s Locomotor activity (total 24 h distance) in 12-month-old KO vs. WT mice: females (KO: n = 7; WT: n = 5) and males (KO: n = 6; WT: n = 4). t Body weight of WT (n = 5 mice per sex group) and Ncoa1 KO mice (females n = 7 and males n = 5). Values are mean ±SEM. Two-way ANOVA with Fisher’s LSD test was applied (panels d, f); unpaired two-sided t-test used for panels e, and g–t. Source data provided as a Source Data file.