Figure 1.

PINK1 and Parkin are phosphorylated after a decrease in ΔΨm in mouse primary neurons. Neurons were infected with lentivirus encoding PINK1-Flag (A), wild-type HA-Parkin (B) or HA-Parkin with either the S65A or S65E mutation (C). Cells were treated with the mitochondrial uncoupler CCCP (30 μm) for 1–3 h and subjected to SDS-PAGE in the absence or presence of 50 μm phos-tag. Note that mobility does not reflect the molecular weight of proteins in phos-tag PAGE (Kinoshita et al. ¹⁵), and thus, molecular weight markers are not shown in the bottom gels. The red and black asterisks in (C) indicate phosphorylation of Parkin at Ser65 and an additional minor phosphorylation site, respectively.