Fig. 4.
Exercise reduced mitochondrial injury and restores mitophagy in senescent CMECs. (A) Mitochondrial O2•− was detected by MitoSOX staining (Top). Fluorescence images of JC-1 monomers (Green) showing hyperpolarized mitochondria and JC-1 aggregates (red) showing normal mitochondria in CMECs from different groups (Bottom). Scale bar, 50 μm. (B) Quantification of relative levels of mitochondrial ROS and potential (Red/Green fluorescence ratio). (C) and (D) CMECs transfected with mt-Keima followed by FCCP (10 μM) treatment showing the mitochondrial network in green and mitophagic vesicles in red. Scale bar, 25 μm. (E) and (F) Western blotting showing levels of LC3 and p62 in mitochondrial fraction and cytosolic fraction of CMECs. (G) and (H) Western blotting showing the mitophagy receptor FUNDC1 level in whole cell lysates of CMECs. n = 4 independent cell isolations per group. **P < 0.01 versus young mice. #P < 0.05, ##P < 0.01 versus aged mice.