Figure 2.

BCAS3 and C16orf70 are recruited to the phagophore assembly site upon mitophagy in a PRKN-PINK1 dependent manner. (A and B) HeLa cells stably expressing 3FLAG-BCAS3 or C16orf70-3FLAG with (A) or without YFP-PRKN (B) were treated with DMSO or valinomycin for 3 h and then immunostained. Bars: 20 µm. (A) Magnified images are shown in the rightmost panels. (C) HCT116 cells stably expressing GST-PRKN were treated with DMSO or valinomycin for 3 h and then immunostained. Endogenous BCAS3 was detected with an anti-BCAS3 antibody. Bars: 10 µm. (D) Total cell lysates in (A) were analyzed by western blotting. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. Orange arrowheads denote ubiquitinated TOMM20