Figure 2.

CCCP-induced mitophagy requires parkin and PINK1 expression. (A) SH-SY5Y cells containing an empty vector (SH) or Park OE cells were treated with vehicle [0.05% (v/v) ethanol] or 10 μm CCCP, lysed and CS activity assessed. Data are expressed as % CS activity of vehicle-treated cells (mean ± SEM; n = 5). *P < 0.05 versus vehicle and **P < 0.01 versus vehicle. (B) Cells were treated with vehicle or CCCP for 24 h, lysed and mitochondrial protein expression (TFAM, MTCOII) assessed by western blot. Equal loading was assessed by probing for GAPDH. Blot representative of four separate experiments. (C) Normal SH-SY5Y cells (control), parkin KD cells or Park OE cells were treated with vehicle or CCCP for 16 h, lysed and CS activity measured. Data are expressed as nmol/min/mg protein (mean ± SEM; n = 5). *P < 0.05 versus vehicle and **P < 0.01 versus vehicle. (D) SH-SY5Y cells or Park OE cells were treated with PINK1 siRNA or control siRNA for 72 h. For the last 16 h, cells were treated with vehicle or CCCP. CS activity was then measured. Data are expressed as nmol/min/mg protein (mean ± SEM; n = 5). *P < 0.05 versus control siRNA + CCCP and **P < 0.01 versus control siRNA + CCCP.