Metabolic index assessment, morphology characterization and fatty acid metabolism analysis of WT and STING KO mice. (A and B) Triglyceride (TG) content of livers (A) and gastrocnemius muscles (B) were measured in 8–12 wk old WT and STING KO mice (n = 9) before and after exercise. Serum biochemistry indexes were assessed in 8–12 wk old WT and STING KO mice before and after exercise. NS, not significant. Unpaired two-tailed T test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (C–H) Serum TG (C; n = 9), serum FFA (D; n = 9), serum lactate (E; n = 9), serum glycerol (F; n = 9), serum cholesterol (G; n = 9), serum insulin (H; n = 9) were measured. NS, not significant; *, P ≤ 0.05. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (I–L) Body weight (I; n = 18,19), total fat mass (J; n = 7), total lean mass (K; n = 7) and food intake (L; n = 11), of 8–12 wk old mice with indicate genotypes were measured. Data are represented as mean ± SEM. ST, STING; NE, non-exercise; EX, exercise. NS, not significant; *, P ≤ 0.05; **, P ≤ 0.01. Unpaired two-tailed t test for comparison of two groups. (M and N) Lipolysis activity in WT mice and STING KO mice. Ex vivo lipolysis assay for eWAT explants from 8–12 wk old WT and STING KO mice before and after exercise (n = 5). Free fatty acid (FFA; M) and glycerol release (N) were measured. Data are represented as mean ± SEM. NS, not significant; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (O) Representative images of Oil-Red staining in mice liver with indicated genotypes before and after exercise. Scale bar, 50 µm. (P) The relative metabolite abundance of long-chain acyl-carnitines in WT and STING KO mice gastrocnemius muscles upon exercise or non-exercise (n = 5). NS, not significant. (Q and R) The translocation of CD36 to plasma membrane in STING KO mice after exercise. Representative images (Q) and quantification (R) of CD36 co-localization of Dystrophin in mice tibialis anterior muscles (n = 4) with indicated genotypes before and after exercise. Data are represented as mean ± SEM. An area of 441 mm2 per mouse was evaluated. ST, STING; NE, non-exercise; EX, exercise. NS, not significant, *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. Scale bar, 20 μm.
Figure S1.

Metabolic index assessment, morphology characterization and fatty acid metabolism analysis of WT and STING KO mice. (A and B) Triglyceride (TG) content of livers (A) and gastrocnemius muscles (B) were measured in 8–12 wk old WT and STING KO mice (n = 9) before and after exercise. Serum biochemistry indexes were assessed in 8–12 wk old WT and STING KO mice before and after exercise. NS, not significant. Unpaired two-tailed T test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (C–H) Serum TG (C; n = 9), serum FFA (D; n = 9), serum lactate (E; n = 9), serum glycerol (F; n = 9), serum cholesterol (G; n = 9), serum insulin (H; n = 9) were measured. NS, not significant; *, P ≤ 0.05. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (I–L) Body weight (I; n = 18,19), total fat mass (J; n = 7), total lean mass (K; n = 7) and food intake (L; n = 11), of 8–12 wk old mice with indicate genotypes were measured. Data are represented as mean ± SEM. ST, STING; NE, non-exercise; EX, exercise. NS, not significant; *, P ≤ 0.05; **, P ≤ 0.01. Unpaired two-tailed t test for comparison of two groups. (M and N) Lipolysis activity in WT mice and STING KO mice. Ex vivo lipolysis assay for eWAT explants from 8–12 wk old WT and STING KO mice before and after exercise (n = 5). Free fatty acid (FFA; M) and glycerol release (N) were measured. Data are represented as mean ± SEM. NS, not significant; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. (O) Representative images of Oil-Red staining in mice liver with indicated genotypes before and after exercise. Scale bar, 50 µm. (P) The relative metabolite abundance of long-chain acyl-carnitines in WT and STING KO mice gastrocnemius muscles upon exercise or non-exercise (n = 5). NS, not significant. (Q and R) The translocation of CD36 to plasma membrane in STING KO mice after exercise. Representative images (Q) and quantification (R) of CD36 co-localization of Dystrophin in mice tibialis anterior muscles (n = 4) with indicated genotypes before and after exercise. Data are represented as mean ± SEM. An area of 441 mm2 per mouse was evaluated. ST, STING; NE, non-exercise; EX, exercise. NS, not significant, *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. Scale bar, 20 μm.

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