Figure 3: ETC-1002 mediates effects on lipid metabolism independently of AMPK β1.

Apoe^−/− and Apoe^−/−/Ampkβ1^−/− (DKO) mice were fed a HFHC diet for 12 weeks with or without ETC-1002 targeted to achieve a 30 mg kg⁻¹ per day dose and (a) total and phosphorylated (T172) AMPKα and total and phosphorylated (S79) ACC measured in livers from Apoe^−/− (n=4) and DKO (n=3). (b) Slc27a2 (ACSVL1) expression in liver and tibialis anterior from untreated Apoe^−/− mice (n=6). (c) Representative fast performance liquid chromatography tracings from chow- and HFHC diet-fed±ETC-1002 Apoe^−/− and DKO mice. Plasma (d) very-low-density lipoprotein (VLDL), (e) LDL, (f) HDL (n=4 for chow-fed mice and 6 for remaining treatment groups) and (g) total cholesterol (n=10), liver (h) cholesterol and (i) triglycerides (n=4 for chow-fed and 6 for remaining treatment groups) determined at the end of study. (j) RER was measured over 24 h following 10 weeks on diet and mean RER during (k) dark (19:00–07:00) and (l) light (07:00–19:00) cycles calculated. (m) Total lipid synthesis in primary hepatocytes isolated from WT and AMPK β1 KO mice treated for 4 h with the indicated concentration of ETC-1002, A-769622 or vehicle (CON) in the presence of ³H-acetate (n=3–8 independent experiments performed in duplicate). (n) Srebf2, Ldlr, Pcsk9, Hmgr, Acly, Srebf1c and Slc27a2 gene expression (n=4 chow-fed and 5 for remaining treatment groups) determined at the end of study. (o) Representative images for immunohistochemistry staining of LDLR in frozen liver sections from ETC-1002-treated Apoe^−/− and DKO mice (20 × ); scale bar, 100 μM. Data are expressed as mean±s.e.m. Multiple comparisons were made using an *one-way (within Apoe^−/− treatment groups) or #two-way ANOVA (between Apoe^−/− and DKO treatment groups); Bonferroni’s multiple comparisons test; *and ^#P<0.05.