Fig. 4

(A-C) pCDNA6.0-His-Gag plasmid was transfected into HEK293T cells, and mitochondria were extracted after 48 h. Changes in the protein levels of Parkin and PINK1 in the mitochondria were detected by Western blot and statistically analyzed (n = 3, *p < 0.05, **p < 0.01, ns p > 0.05). (D) shRNA targeting endogenous Parkin was transfected into HEK293T cells for 24 h, followed by transfection with pCDNA6.0-His-Gag plasmid for an additional 48 h. Cell lysates were collected to extract protein, and Western blot analysis was performed to detect changes in Tom20 and Cox4. (E-F) Statistical analysis of Tom20 and Cox4 protein expression changes (n = 3, *p < 0.05, **p < 0.01, ns p > 0.05). (G) Following the transfection of shPark2 to knock down endogenous Parkin in HEK293T cells, mCherry-GFP-LC3 and pCDNA6.0-His-Gag plasmid were cotransfected. The co-localization of LC3 and Tom20 was analyzed using confocal microscopy. (H) Statistical analysis of the co-localization of LC3 and Tom20. Scale bar = 10 μm. (***p < 0.001)