Loss of hepatic dynamin-related protein 1 (DRP1) exacerbates EtOH-impaired mitophagy in hepatocytes. (A) Primary hepatocytes from wild-type (WT) and liver-specific DRP1 knockout (L-DRP1 KO) mice were infected with Ad-Cox8-EGFP-mCherry overnight. Cells were treated with or without ethanol (80 mM) for 6 hours, followed by fluorescence microscopy. (B) The number of red puncta in each cell was quantified (n = 40–60 of 3 mice). (C) Male 2–3-month-old L-DRP1 KO and matched littermates were subjected to Gao-binge alcohol feeding. Ad-Cox8-EGFP-mCherry (5 × 108 PFU per mouse, intravenously) were injected on the fifth day. Representative confocal microscopy images of cryo-liver sections are shown. (D) The number of red puncta in each cell was quantified. Data are presented as means ± SD (n = 3 mice). (E) Total liver lysate of WT and L-DRP1 KO mice fed with control or EtOH diet were subjected to western blot analysis. (F) Densitometry analysis of (E). Data are presented as means ± SEM (n = 3–4). *p < 0.05, **p < 0.01, ***p < 0.001; one-way analysis of variance analysis with Bonferroni’s post hoc test. EGFP, enhanced green fluorescent protein; EtOH, ethanol.