Fig. 5

The effect of AA on mitophagy, mitochondrial turnover and cell apoptosis. H9C2 cells were treated as described in Fig. 2. Western blot was used to measure the expression of mitophagy associated proteins, mitochondrial renewal proteins and apoptosis-related factors. Real-time PCR was used to detect the level of mitochondria and genomic DNA. A–D quantification of mitophagy associated proteins LC3II/LC3I (A), p62 (B), PINK1 (C) and parkin (D); E, F quantification of mitochondrial renewal proteins Drp1 (E) and FIS1 (F); H–J, quantification of apoptosis-related factors cytosolic cytochrome c (cyt c) (H), mitochondrial cyt c (mito-cyt c) (I), and cleaved caspase-3 (J). GAPDH and VDAC1 were used as internal protein loading controls respectively for cytosolic and mitochondrial protein. Optical densities of the protein bands were quantitatively analyzed with Sigma Scan Pro 5 and normalized with loading control GAPDH or VDAC1. G mitochondrial to genomic DNA. Results are expressed as the mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001. Statistical analysis was carried out by a one-way ANOVA analysis