Autophagy factors colocalize with ΔOTC foci in a PINK1-dependent manner. (A) Tet ON: ΔOTC-expressing HeLa cells with or without a PINK1 KO background and with or without YFP-Parkin expression expressing mCherry-LC3 (LC3; red) or mCherry-optineurin (OPTN; red) were treated with DOX for 48 h, fixed, stained for TOM20 (blue), and imaged. (B) Tet-ON: ΔOTC-expressing cells with YFP-Parkin expression with or without mCherry-LC3 or mCherry-optineurin (red) expression were treated with DOX for 48 h and processed for indirect immunofluorescence microscopy with an antibody against OTC (green) and ubiquitin (Ub; red).Bar, 10 µm. (C) Tet-ON: ΔOTC-expressing cells without YFP-Parkin expression with or without mCherry-LC3 or mCherry-optineurin (red) expression were treated with DOX for 48 h and processed for indirect immunofluorescence microscopy with an antibody against OTC (green) and ubiquitin (red). Bars, 10 µm. (D) Quantification of the percentage of foci of LC3, optineurin, or ubiquitin that colocalize with ΔOTC in the cells with and without Parkin expression as described in B and C. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Error bars indicate SD.