FIGURE 4. HSG4112 regulates metabolic gene expression, influences AMPK protein level, and has no effect on UCP-1 level. (A-D) Relative gene expression was measured using qRT-PCR (n = 4) on terminal (A) gastrocnemius muscle, (B) liver, (C) hypothalamus, and (D) interscapular adipose tissue (BAT) collected from the same mice from Figure 2. Data represent mean ± SEM. *P < 0.05, **P < 0.01 vs. Vehicle group,#P < 0.05,##P < 0.01 vs. Normal group; one-way ANOVA with Tukey’s multiple comparison test. (E, F) HFD-mice were treated for 11 days before sacrifice (n = 4 for AMPK groups, 3 for UCP1 groups). (E) Western blot of phosphorylated hypothalamic AMPK ɑ1 and ɑ2 and their quantification. *P < 0.05, two-way ANOVA with Sidak’s multiple comparison test; (F) Western blot of BAT UCP1 and its quantification. n.s. = not significant; Student’s t-test.