Extended Data Fig. 7: IL1R1 deficiency or IL1R1 antagonist treatment reduces MAP kinase phosphorylation.
From: IL1R1 is required for celastrol’s leptin-sensitization and antiobesity effects
a–d, Ad libitum–fed Il1r1+/+ mice and food-restricted (FR) Il1r1–/– mice (~0.5 g of food per day) were treated with vehicle (Veh) or celastrol (Cel; 100 μg/kg body weight, i.p., daily) for 7 d and the phosphorylation status of hypothalamic or hepatic MAP kinases (p38, Erk1/2) was analyzed by western blot. a, Representative immunoblots for phoshorylated p38 MAP kinase (p-p38) and total p38 MAP kinase (p38) in the hypothalamus of Il1r1+/+ and FR Il1r1–/– mice. b, Ratio of quantified p-p38 density to p38 density. n = 3 mice for each group. Il1r1+/+–Veh vs. Il1r1–/––Veh, P < 0.0001; Il1r1+/+–Cel vs. Il1r1–/––Cel, P < 0.0001; Il1r1–/––Veh vs. Il1r1–/––Cel, P > 0.99. c, Representative immunoblots for phosphorylated MAP kinase (p-Erk1/2) and Erk1/2 MAP kinase in the liver of Il1r1+/+ and FR Il1r1–/– mice. d, Ratio of quantified p-Erk1/2 density to Erk1/2 density. n = 3 mice for each group. Il1r1+/+–Veh vs. Il1r1+/+–Cel, P = 0.06; Il1r1+/+–Cel vs. Il1r1–/––Cel, P = 0.01; Il1r1–/––Veh vs. Il1r1–/––Cel, P > 0.99. e,f, DIO mice were administered the IL1R1 antagonist anakinra (Ank; 5 μg/mouse per day) through intracerebroventricular injection into the lateral ventricle in combination with celastrol (100 μg/kg body weight, i.p., daily) for 7 d. e, Representative immunoblots for p-p38 and total p38 in the hypothalamus. f, Ratio of quantified p-p38 density to p38 density. n = 5 mice for each group. P = 0.03. g,h, DIO mice were treated with IL1R1 antagonist (Ank; 30 mg/kg body weight, i.p., twice a day) in combination with celastrol (100 μg/kg body weight, i.p., daily) for 7 d. g, Representative immunoblots for p-Erk1/2 and total Erk1/2 in the liver. h, Ratio of quantified p-Erk1/2 density to Erk1/2 density. n = 6 mice for each group. Sal + Veh vs. Sal + Cel, P = 0.0004; Ank + Veh vs. Ank + Cel, P > 0.99; Sal + Cel vs. Ank + Cel, P = 0.0004. Values indicate averages ± s.e.m. P values were determined by two-way ANOVA with Bonferroni’s multiple-comparisons test (b,d,h) or two-tailed Student’s t test (f). *P < 0.05, ***P < 0.001; n.s., not significant (P > 0.05).
