Fig. 3

Parkin is required for the activation of PD-induced mitophagy both in vivo and in vitro. a Wild-type or Parkin−/− mice were subjected to ARDS, and then treated with PD or a vehicle for 12 h. Mitochondrial markers, COX4I1 and TIM23, as well as mitochondrial biogenesis marker proteins, PGC-1α and mtTFA, were measured by western blot. b Quantification analysis of COX4I1 levels. c Quantification analysis of PGC-1α levels. d Quantification analysis of TIM23 levels. e Quantification analysis of mt-TFA levels. f Beas-2B cells were transfected with Parkin siRNA (a scrambled siRNA was used as a control) and mt-Keima-COX8 for 24 h. The knockdown of Parkin expression was confirmed by western blot. The cells were subsequently exposed to LPS (0.5 mM) and treated with either PD (50 μM) or a vehicle for 6 h. Laser confocal-scanning microscopy was performed to observe mitochondria-localised Keima (green) and the acidic Keima puncta (red) delivered within autolysosomes. Scale bar: 10 μm. g Quantification of the acidic Keima per cell. Data are presented as the mean ± SD (n = 6 in each group). *P < 0.05 vs. the indicated groups; #P > 0.05 vs. the indicated groups