Fig. 6: Dio3os activation in brown fat (BAT) protects maternal obesity (MO) female offspring from western-diet-induced obesity and fatty liver at 22 °C.

a 8-week-old female offspring born from normal and MO dams were injected with AAV8-scramble-CMV or AAV8-Dio3os-CMV viral particles (5 × 10¹¹ vg per mouse) into interscapular BAT, respectively. Control and MO offspring at the same age were treated by saline injection into interscapular BAT as negative controls. After injection, offspring were fed HFD for 5 weeks at 22 °C. b mRNA expression of Dio3os in offspring BAT at 5-week post injection (n = 5). c, d body weight (c) and BAT mass (d) in offspring post-injection (n = 5). e UCP-1 immunostaining in offspring BAT at 5-wk post injection. H&E staining of BAT, subcutaneous, visceral white fat, and liver. Scale bar 250 µm (immunostaining) and 200 µm (H&E) (n = 5). f Immunoblotting measuring protein contents of PRDM16, PGC-1a, and UCP-1 in offspring BAT at 5-wk post-injection (n = 5). β-tubulin was the loading control. g Inguinal, visceral fat and liver mass in offspring at 5-wk post injection (n = 5). h Blood glucose in offspring after intraperitoneal glucose injection for measuring glucose tolerance (n = 5). i Offspring energy expenditure at 5-week post injection (n = 5). Metabolic data were regressed to body mass according to guidelines of ANCOVA NIDDK MCCP tool. Data are presented as mean ± s.e.m. Two-way ANOVA with Bonferroni post hoc was used in data analysis.