FIGURE 8.
Acute knockdown of Parkin in mouse livers impaired with APAP-induced mitophagy and exacerbated APAP-induced liver injury. A, male C57Bl/6J mice were treated either with Ad-Neg or Ad-Parkin shRNA (i.v., 1 × 109 pfu per mouse) for 4 days. Total liver, pancreas, and skeletal muscle lysates were subjected to Western blot analysis. Lysate from a Parkin KO mouse liver was used a negative control. Mice were further treated with APAP (500 mg/kg) or saline for another 6 h. B, blood samples were used to measure serum ALT levels. Data shown are means ± S.E. (n = 4; *, p < 0.05). C, representative H&E images are shown (×200 magnification). Dotted line circled areas denote centrilobular necrosis. D and E, total liver lysates were subjected to Western blot analysis. Densitometry quantification of Western blots for p-JNK1 and p-JNK2 (n = 3 per group). Results were normalized to β-actin. F, representative EM images are shown. An enlarged image of a mitophagosome showing an enveloped mitochondrion and an enlarged image of an autophagosome showing enveloped cytosolic proteins are from the dotted line boxed areas (arrow = mitophagosomes, M = mitochondria, and N = nucleus). G and H, quantification of EM images for mitophagosomes and total autophagosomes. Data shown are means ± S.E. (n ≥ 20 images quantified per group; *, p < 0.05).