Fig. 5

Adipose-selective expression of H3.3K36M depletes H3K36 methylation and reprograms BAT. a Schematic of transgenic (A-K36M) construct. Expression of FLAG-tagged H3.3K36M is under the control of the 7.9 kb adipose tissue-selective Fabp4 promoter. b Tissue distribution of FLAG-tagged H3.3K36M in A-K36M mice was determined by Western blot. All data in this figure were from 8-week-old to 10-week-old male mice fed with regular diet. c Western blot in BAT of control (Ctrl) and A-K36M mice. d Representative picture of Ctrl and A-K36M mice (left). Fat mass, lean mass, and total body weight of Ctrl and A-K36M mice (n = 6 per group) were measured by MRI (right). e Representative pictures of interscapular WAT (iWAT), BAT, retroperitoneal WAT (rWAT), inguinal WAT (ing-WAT), epididymal WAT (epi-WAT), and liver. f Average tissue weights in Ctrl and A-K36M mice (n = 6 per group) are presented as % of body weight. g Enlarged images (upper panels) and H&E staining (lower panels) of BAT isolated from Ctrl and A-K36M mice. Scale bar = 100 µm. h qRT-PCR analysis in BAT of Ctrl and A-K36M mice (n = 6 per group). i Western blot of PPARγ, C/EBPα, and Ucp1 in BAT isolated from Ctrl and A-K36M mice. j–l BAT and epi-WAT were collected from A-K36M and Ctrl mice for RNA-Seq. j Heatmaps of BAT-enriched or epi-WAT-enriched genes. Column 1, log fold change between RPKM of BAT vs. epi-WAT in Ctrl mice. Top 1000 WAT-selective or top 1000 BAT-selective genes were analyzed. Columns 2–3, log fold change between RPKM of A-K36M vs. Ctrl mice (A-K36M/Ctrl) in BAT (column 2) or epi-WAT (column 3). k, l RPKM values of WAT-selective (k) or BAT-selective (l) genes. m, n Cold tolerance test. Ctrl and A-K36M mice (n = 6 per group) were housed at room temperature (22 °C) or in cold (4 °C) for 2 h. m Body temperatures. n qRT-PCR of Ucp1 and Dio2 in BAT. All values in d, f, h and m, n are presented as mean ± SEM. Statistical comparison between groups was performed using Student’s t test. *p < 0.05, **p < 0.01, and ***p < 0.005