Fig. 5: H1.2 regulates Il10rα and thermogenesis in beige adipocytes in vitro.
From: Linker histone variant H1.2 is a brake on white adipose tissue browning
a, b H1.2 mRNA level (a) and representative Oil red O staining (b) in differentiated beige adipocytes isolated from H1.2flox/flox mice treated with adenovirus-packed Cre recombinase ( + Cre) or vehicles (-Cre) (n = 3, repeated three times independently with similar results obtained). Scale bar = 50 μm. c–f mRNA levels of genes related to adipogenesis (c), thermogenesis (d) and lipid β-oxidation (e), and protein levels of H1.2 and Ucp1 (f) of differentiated beige adipocytes isolated from H1.2flox/flox mice treated with or without Cre (n = 3). g, h Il10rα mRNA level (g) and representative Il10rα and Ucp1 protein levels (h) in differentiated beige adipocytes isolated from H1.2flox/flox mice, with or without Cre/10 μM isoprenaline (Iso) treatments (n = 3 in g; n = 2 in h; repeated three times independently with similar results obtained). i qPCR of thermogenic genes of differentiated beige adipocytes isolated from H1.2flox/flox mice, with or without Cre/Il10 treatments (n = 3). PH1.2 (-cre+Il10-0 vs -cre+Il10-50) = 0.8879, PH1.2 (-cre+Il10-0 vs -cre+Il10-100) = 0.1621, PH1.2 (-cre+Il10-0 vs +cre+Il10-0) < 0.0001; PIl10ra (-cre+Il10-0 vs -cre+Il10-50) = 0.2056, PIl10ra (-cre+Il10-0 vs -cre+Il10-100) = 0.1964, PIl10ra (-cre+Il10-0 vs +cre+Il10-0) = 0.0433; PUcp1(-cre+Il10-0 vs -cre+Il10-50) = 0.0105, PUcp1 (-cre+Il10-0 vs -cre+Il10-100) = 0.0007, PUcp1 (-cre+Il10-0 vs +cre+Il10-0) < 0.0001, PUcp1 (+cre+Il10-0 vs +cre+Il10-50) = 0.4609, PUcp1 (+cre+Il10-0 vs +cre+Il10-100) = 0.0639; PDio2(-cre+Il10-0 vs -cre+Il10-50) = 0.0005, PDio2 (-cre+Il10-0 vs -cre+Il10-100) = 0.0002, PDio2 (-cre+Il10-0 vs +cre+Il10-0) < 0.0001, PDio2 (+cre+Il10-0 vs +cre+Il10-50) = 0.1604, PDio2 (+cre+Il10-0 vs +cre+Il10-100) = 0.2028; PCidea (-cre+Il10-0 vs -cre+Il10-50) = 0.3045, PCidea (-cre+Il10-0 vs -cre+Il10-100) = 0.0276, PCidea (-cre+Il10-0 vs +cre+Il10-0) = 0.0005, PCidea (+cre+Il10-0 vs +cre+Il10-50) = 0.3547, PCidea (+cre+Il10-0 vs +cre+Il10-100) = 0.0737; PCox8b (-cre+Il10-0 vs -cre+Il10-50) = 0.0296, PCox8b (-cre+Il10-0 vs -cre+Il10-100) = 0.0236, PCox8b (-cre+Il10-0 vs +cre+Il10-0) = 0.0552, PCox8b (+cre+Il10-0 vs +cre+Il10-50) = 0.6737, PCox8b (+cre+Il10-0 vs +cre+Il10-100) = 0.7483. j Ucp1 and Il10rα levels of differentiated beige adipocytes isolated from H1.2flox/flox mice, with or without Cre/Il10 treatments. k Il10rα level in differentiated beige adipocytes isolated from H1.2AKO mice, with or without AAV-Il10rα treatments (n = 3; repeated three times independently with similar results obtained). l Ucp1 and Dio2 levels in differentiated beige adipocytes isolated from H1.2AKO mice, with or without AAV-Il10rα/Il10 treatments (n = 3). PUcp1(AAV-Vehicle+Il10-0 vs AAV-Vehicle+Il10-50) = 0.9997, PUcp1 (AAV-Vehicle+Il10-0 vs AAV-Vehicle+Il10-100) = 0.9839, PUcp1 (AAV-Vehicle+Il10-0 vs AAV-Il10rα+Il10-0) = 0.0043, PUcp1 (AAV-Il10rα+Il10-0 vs AAV-Il10rα+Il10-50) = 0.0467, PUcp1 (AAV-Il10rα+Il10-0 vs AAV-Il10rα+Il10-100) = 0.0019, PUcp1 (AAV-Il10rα+Il10-50 vs AAV-Il10rα+Il10-100) = 0.0141; PDio2 (AAV-Vehicle+Il10-0 vs AAV-Vehicle+Il10-50) = 0.8793, PDio2 (AAV-Vehicle+Il10-0 vs AAV-Vehicle+Il10-100) = 0.9737, PDio2 (AAV-Vehicle+Il10-0 vs AAV-Il10rα+Il10-0) = 0.0104, PDio2 (AAV-Il10rα+Il10-0 vs AAV-Il10rα+Il10-50) = 0.0451, PDio2 (AAV-Il10rα+Il10-0 vs AAV-Il10rα+Il10-100) = 0.0228, PDio2 (AAV-Il10rα+Il10-50 vs AAV-Il10rα+Il10-100) = 0.9999. m Ucp1 and Il10rα levels of differentiated beige adipocytes isolated from H1.2AKO mice, with or without AAV-Il10rα/Il10 treatments (n = 2; repeated three times independently with similar results). Data are mean ± S.D. Unpaired two-tailed Student’s t test in a, c–f, k; one-way ANOVA with Tukey’s multiple comparisons test in g, i and l. *P < 0.05, **P < 0.01; ns not significant. Source data and exact P values are provided in a Source data file.
