Figure 6.

p32 regulates autophagy and mitophagy via interacting with ULK1. (a) Hela cells stably reconstituted with lentivirus bearing the indicated p32 cDNA were infected with p32-shRNA lentiviruses. Lysates were immunoblotted with the indicated antibodies. (b) H1299-GFP-LC3 cells stably expressing the indicated shRNA-resistant forms of p32 were infected with p32-shRNA lentiviruses. Cells were then treated with EBSS medium for 6 h. GFP-LC3 positive autophagosomes were examined by confocal microscopy. The scale bar represents 20 μM (left panel). Histogram represents the percentage of cells containing positive LC3 punctas (right panel). The error bars represent the mean±S.D. of five random areas. **P<0.01. (c) Hela cells stably expressing GFP-Parkin and the indicated shRNA-resistant forms of p32 were infected with p32-shRNA lentiviruses. Cells then were treated with CCCP (10 μM) for 48 h. Cells were immunostained with anti-TOM20 antibody (upper) and scored for the percentage of cells expressing GFP-parkin with TOM20 staining, which was shown as mean±S.D. of five random areas (lower). The scale bar represents 10 μM. ***P<0.001