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. 2020 May 13;6(20):eaay6191. doi: 10.1126/sciadv.aay6191

Fig. 3. Adipocyte sST2 expression and secretion are stimulated by TNFα, which is antagonized by Zbtb7b.

Fig. 3

(A) Correlation between eWAT sST2 gene expression and body weight (left) and Ccl2 expression (right). (B) qPCR analysis of sST2 mRNA expression in differentiated C3H10T1/2 adipocytes treated with vehicle (Veh), IL-1β (10 ng/ml), TNFα (10 ng/ml), or IFN-γ (10 ng/ml) for 24 hours (left) or indicated doses of TNFα (right). Data represent means ± SD (n = 3). **P < 0.01, ***P < 0.001, versus Veh, one-way ANOVA. (C) The concentrations of sST2 in CM from C3H10T1/2 adipocytes (left) or eWAT explant culture (right) treated with Veh or TNFα (10 ng/ml) for 24 hours. Data represent means ± SD (n = 3). ***P < 0.001, Veh versus TNFα, two-tailed unpaired Student’s t test. (D) sST2 mRNA expression (left) and concentrations (right) in adipocytes treated with TNFα (10 ng/ml) with or without NF-κB inhibitor VIII (VIII; 4 μM) for 24 hours. Data represent means ± SD (n = 3). ***P < 0.001, Veh versus VIII, two-way ANOVA. Data in (B) to (D) are representative of three independent experiments. (E) Correlation between eWAT mRNA expression of Zbtb7b and bodyweight (left) and sST2 (right) in HFD-fed mice. (F) qPCR analysis of Zbtb7b expression in SVF and adipocytes isolated from chow-fed (filled, n = 3) or HFD-fed (open, n = 3) mouse eWAT. Data represent means ± SEM. ***P < 0.001, SVF versus Adp, two-way ANOVA. (G) qPCR analysis of Zbtb7b expression in eWAT from WT (filled, n = 5) and ob/ob (open, n = 3) or WT (filled, n = 4) and db/db (open, n = 4) mice. Data represent means ± SEM. *P < 0.05, **P < 0.01, lean versus obese, two-tailed unpaired Student’s t test. (H and I) qPCR analysis of Zbtb7b expression in differentiated C3H10T1/2 adipocytes (H) and explant fat culture (I) treated with 10 ng/ml TNFα for 24 hours. Data in (H) and (I) represent means ± SEM. *P < 0.05, Veh versus TNFα, two-tailed unpaired Student’s t test. (J) sST2 mRNA expression (left) and concentrations in CM (right) from C3H10T1/2 adipocytes expressing vector (Vec) or Zbtb7b following treatment with TNFα (10 ng/ml) for 24 hours. Data represent means ± SD (n = 3). ***P < 0.001, Vec versus Zbtb7b, two-way ANOVA. Data in (F) to (J) are representative of three independent experiments.