Figure 5.

Acute exercise enhances mitochondrial ATF5 in WT mice, while there is reduced basal mitochondrial Parkin, attenuated exercise-induced mitochondrial LC3-II and enhanced stress signaling with exercise in ATF5 KO animals. A) Representative blots of ATF5 in nuclear, cytosolic and SS and IMF mitochondrial fractions from CON or EX animals subjected to acute exercise (continuous and exhaustive). The blot from nuclear and cytosolic samples is shown with the corresponding Ponceau stain, while that from mitochondrial samples is shown with VDAC. B) Graphical representation of ATF5 protein in different cellular fractions and conditions. Blots corrected for Ponceau or VDAC and displayed as a fold change (EX/CON) (n = 6–9). Protein loaded for each fraction is as follows: Nuclear, 60 $\mu$g; Cytosolic, 10 $\mu$g; Mitochondrial, 25 $\mu$g. Representative blots of C) whole muscle p-JNK, p-eIF2⍺, t-JNK and t-eIF2⍺ and D) autophagy proteins LC3-I, LC3-II and Parkin in whole muscle and SS and IMF mitochondrial samples in CON and EX animals subjected to acute exercise (continuous and exhaustive). Blots are shown with corresponding Ponceau stains or VDAC blots. E) Quantifications of phosphorylated/total for JNK and eIF2⍺ corrected for Ponceau (n = 5–12). F) Quantification of LC3 in whole muscle represented as the LC3-II/I ratio, and LC3-II and Parkin in SS and IMF mitochondria (n = 6–11). $\Psi$P < 0.05, $\Psi \Psi$P < 0.01 main effect of exercise; #P < 0.05, ##P < 0.01 main effect of genotype; $†$P < 0.05 interaction effect of exercise and genotype; ∗P < 0.05, ∗∗P < 0.01Bonferroni post-hoc analysis, CON vs EX of same genotype unless otherwise indicated. A. U., arbitrary units; C, cytosolic; CON, control; EX, exercised; KO, knockout; N, nuclear; p, phosphorylated; S, standard; t, total; WT, wild-type. Lines in blot indicate where two different areas were spliced together from the same blot.