Extended Data Fig. 3: BAT-derived Nrg4 improved metabolic profiles, alleviated endothelial dysfunction and inflammation or adhesion response in mice.
(A) The Nrg4 protein level in BAT, eWAT, iWAT and liver of mice in KO-AAV (Nrg4) groups (n = 3). (B) Results of GTT (n = 6) and the AUC for GTT (*P < 0.001, n = 6). (C) Normalized the AUC for ITT as a percentage of fasting glucose and the AUC for it (*P < 0.001, n = 6). (D) The energy expenditure of mice aged 18 weeks in different groups (n = 6, *P < 0.001). (E) The blood Nrg4 level in mice of different groups (n = 6). (F) The Ach induced aortic vasodilatation of mice in different groups (n = 6). (G) The experiment schedule for the effects of Nrg4 overexpression in BAT in situ in both AKO and DKO mice (6 mice in each group). (H) The experiment schedule for BATT in KO recipients (n = 6 in each group). (I) The mRNA levels of adhesion molecules (VCAM-1, ICAM-1, and E-selectin) and inflammation (TNF-α, IL-1β, and IL-6) (n = 6). ((E), (F), (I): *P < 0.01 versus DKO-AAV(Zsgreen), #P < 0.05 versus AKO-AAV(Zsgreen). *P < 0.05 vs. AKO-AAV(Zsgreen); **P < 0.01 vs. AKO-AAV(Zsgreen); ***P < 0.001 vs. AKO-AAV(Zsgreen); #P < 0.05 vs. DKO-AAV(Zsgreen); ##P < 0.01 vs. DKO-AAV(Zsgreen); ###P < 0.001 vs. DKO-AAV(Zsgreen)). (J) The confirmation experiments for circulating expression of Nrg4 in BATT KO recipients. All mice aged 12 weeks were treated with BATT and then fed a WD for 12 weeks (n = 3). (K) The experiment schedule for BATT in AKO and DKO mice (n = 6 in each group). (L) Results of GTT (n = 6). (M) The AUC for GTT (n = 6). (N) Normalization ITT as a percentage of fasting glucose. (O) The AUC for (N) (n = 6). (P) The energy expenditure of mice aged 24 weeks. (n = 6). (L-P: *P < 0.05 vs. BATT(KO); **P < 0.01 vs. BATT(KO); ***P < 0.001 vs. BATT(KO); #P < 0.05 vs. BATT(KO); ##P < 0.01 vs. BATT(KO); ###P < 0.001 vs. BATT(KO)). Statistical significance was calculated using two-sided t-test or one-way ANOVA with Tukey’s multiple-comparison test. Data were shown as mean ± SEM.
