Fig. 1
Ucp1 expression and H3K9me2 levels in adipose tissues and BAT thermogenic function in Jmjd1a-S265AKI/KI mice . a Ucp1 mRNA (n = 3) (left) and protein levels detected by immunoblotting (IB) (right) in BAT, and scWAT of mice exposed to 4 °C for 6 h. b H3K9me2 immunoblotting using purified histones from adipose tissues of mice housed at RT. c H3K9me2 ChIP-qPCR in BAT, and scWAT of mice exposed to 4 °C for 6 h (n = 3). d NE-induced Ucp1 mRNA levels in im-BATs stably expressing WT or indicated mutants of hJMJD1A. The relative quantity in im-BATs expressing WT-JMJD1A on day 8 before NE treatment (0 h) is defined as 1. e Ucp1 mRNA expressions (n = 3) (left) and proteins (right) in BAT, and scWAT from mice exposed to 30 °C or 4 °C for 1 week. Uncropped images of the blots (a, b, e) are shown in Supplementary Fig. 8. f, g H3K9me2 ChIP-qPCR in scWAT from mice placed at 4 °C for 1 week (n = 4) (f) and in im-scWATs during beige adipogenesis (mean ± s.e.m. of three technical replicates) (g). h Bi-phasic Ucp1 expressions during cold exposure in BAT and scWAT. Mice were exposed to 4 °C for the indicated time, and Ucp1 mRNAs in BAT and scWATs were quantified by qPCR (n = 3). The data were converted to copy number per ng of total RNA. i In BAT, Ucp1 locus is in euchromatin (left), while in scWAT, it is in heterochromatin with H3K9me2 (right). In BAT, cold exposure leads to acute induction of Ucp1 mRNA through the mechanisms independent of H3K9me2 demethylation (left). In scWAT, H3K9me2 at Ucp1 gene locus needs to be removed for beige adipogenesis (right bottom). j Cold intolerance in Jmjd1a-S265AKI/KI mice (n = 6 per genotype group). Shown is the body temperature of 9-week-old mice at different times after cold exposure (4 °C). k Impaired NE-induced activation of selective genes in BAT (n = 6 per genotype group). l, m Reduced NE-induced mitochondrial respiration (l) and NE-induced glycerol release (m) in primary brown adipocytes from Jmjd1a-S265AKI/KI mice. Data are mean ± s.e.m. of five technical replicates (l) and three independent experiments (m). Data are mean ± s.e.m. a, c, e, f, j, k Student’s t test (a, e, f, j, l, m) or analysis of variance, followed by Tukey’s post hoc comparison (k) were performed for comparisons. *P < 0.05, **P < 0.01, and ***P < 0.005
