Fig. 6: SAPS3 deficiency-mediated metabolic homeostasis in vivo under HFD is AMPK dependent. | Nature Communications

Fig. 6: SAPS3 deficiency-mediated metabolic homeostasis in vivo under HFD is AMPK dependent.

From: SAPS3 subunit of protein phosphatase 6 is an AMPK inhibitor and controls metabolic homeostasis upon dietary challenge in male mice

Fig. 6

a Experimental strategy for feeding 8 weeks old male LKO mice with HFD (45 kcal% fat) for 16 weeks and then treating mice with compound C for two weeks. b Fat mass composition of FF and LKO male mice under HFD before compound C treatment and after compound C treatment for two weeks. Mean ± s.d., n = 6 mice per group analyzed by two-tailed t-test. Before treatment, FF/LKO, **p = 0.006, **p = 0.0017; After treatment, FF/LKO, *p = 0.033; n.s., not significant.; LKO, before treatment/comC, *p = 0.048. c Liver weights from mice treated with ctrl or compound C. Mean ± s.d., n = 8 mice per group analyzed by two-tailed t-test, ***p = 0.0005; n.s., not significant. d AMPK mediated ACC phosphorylation was tested by Western blotting. The results are representative of two independent experiments. e Representative images of H&E staining of liver slides from FF and LKO mice. Scale bar, 100 µm. f The quantification of the H&E staining images shown in e. Mean ± s.d., n = 6 mice per group analyzed by two-tailed t-test, ****p = 8.7918E-08; n.s., not significant. g Experimental strategy for injecting AAV-AMPK-DN in 6 weeks old male LKO mice and, after two weeks, followed by feeding mice with HFD (45 kcal% fat) for 16 weeks. h The inhibition of AMPK activity by AAV-AMPK-DN was evaluated by the level of phospho-ACC from liver samples after HFD. The results are representative of two independent experiments. i Body weight was measured. Mean ± s.d., n = 6 mice per group analyzed by two-tailed t-test, n.s., not significant; or one-way ANOVA, ****p < 0.0001. j Fat compositions were measured by echo-MRI. Mean ± s.d., n = 6 mice per group analyzed by two-tailed t-test, ****p = 5.9683E-06; n.s., not significant. k Representative pictures of mice and livers from FF and LKO mice. And the liver weights were compared. Mean ± s.d., n = 6 mice per group analyzed by two-tailed t-test, ***p = 0.00018; n.s., not significant. l Representative images of H&E staining of liver slides from FF and LKO mice with or without AAV. Scale bar, 100 µm. CD, AAV (−) n = 3 mice per group; CD, AAV ( + ) n = 4 mice per group; HFD, AAV (−) n = 6 mice per group; HFD, AAV ( + ) n = 6 mice per group. m Glucose tolerance test and insulin tolerance test were performed at the end of the experiment. Mean ± s.d., n = 6 per group analyzed by one-way ANOVA or two-tailed t-test. ****p < 0.0001; ***p = 0.0001; n.s., not significant.

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