Figure 7.

Ang- (1-7) prevents the increment of autophagy in gastrocnemius from DDC-treated mice. C57BL/6J male mice were fed a standard diet (Control) or DDC-supplemented diet (DDC) with or without Ang-(1-7) (through osmotic minipumps). Five animals per group were used for three independent experiments. When the experiment finished (week 6), the gastrocnemius muscle was removed, and RNA and protein extraction were performed. (A) LC3 I and LC3 II protein levels were evaluated by western blot. β-actin was used as the loading control. Molecular weights are depicted in kilodaltons. (B) Densitometric analysis of the band was performed, and the ratio of LC3 II/LC3 I is shown. Values are expressed as fold of induction relative to control and represent the mean ± SEM. RT-qPCR determined LC3B (C), p62 (D), and Beclin-1 (E) gene expression. β-actin was used as a housekeeping gene (qPCR). Values represent the mean ± SEM. (For B, C, D and E: (* p < 0.05 vs. Control, # p < vs. DDC. One-way ANOVA, Tukey’s multiple comparisons test).