Fig. 2.
Effect of a HFD on AMPK phosphorylation and CPT1 abundance in the livers of WT and IL-6–/– mice. (A) Western blot analysis of the hepatic expression of AMPK and p-AMPK in samples from WT and IL-6–/– mice fed STD or HFD. The blot shows results from two or three independent samples from each group. Corresponding expression of actin is shown as a loading control per lane. The figure shows one representative blot from two independent experiments and densitometric values of the p-AMPK/AMPK ratio are shown in the histogram below. The values are presented as the means±s.e.m. (n=4 samples per group), and differences between groups were evaluated using two-way ANOVA analysis and Bonferroni post-hoc tests. *P<0.05 and **P<0.001 denote significant differences compared with the corresponding STD-fed group. ###P<0.001 denotes significant differences between both HFD-fed groups. (B) qPCR analysis determining the Cpt1 gene expression in liver samples from WT and IL-6–/– mice fed STD or HFD. The Cpt1 expression was normalized using Biogazelle's qbasePLUS software with Gapdh and Gusβ as reference genes. The columns represent CNRQ means±s.e.m. (n=4 animals per group) and differences between groups were evaluated using two-way ANOVA analysis and Bonferroni post-hoc tests. ***P<0.001 denotes significant differences compared with the corresponding STD-fed group. ###P<0.001 denotes significant differences between the HFD-fed groups. (C) Western blot showing CPT1 protein expression in WT and IL-6–/– mice fed STD or HFD. Three representative samples from each group are shown. The levels of CPT1 were determined through densitometry corrected for adaptin (CPT1/adaptin), and the corresponding values are shown in the histogram below. The figure shows one representative blot from two independent experiments. The values represent the means±s.e.m. (n=4 animals per group), and comparisons between groups were analyzed using two-way ANOVA and Bonferroni post-hoc tests. **P<0.01 denotes significant differences compared with the corresponding STD-fed group. ##P<0.01 denotes significant differences between the HFD-fed groups.