Figure 3.
Ubiquitination of mitochondrial proteins following CCCP treatment. (A) SH-SY5Y cells containing an empty vector or Park OE cells were treated with vehicle [0.05% (v/v) ethanol] or 10 µm CCCP for 3 h, mitochondria isolated and a western blot performed. The western blot was probed with an antibody against ubiquitin. Equal loading was determined by probing for the 39 kDa subunit of complex I (arrow). Blot representative of four separate experiments. (B) Park OE cells were treated with CCCP for 3 h, mitochondria isolated and probed for VDAC1 or (C) prohibitin (PHB1) by western blot. (D) Park OE cells were treated with CCCP for 3 h and cell lysates probed for Drp1 by western blot. (E) Mitochondria isolated from Park OE cells treated with vehicle or CCCP for 3 h were probed for MFN-1 or (F) MFN-2 by western blot. Post-translationally modified proteins are indicated by arrowheads. Non-specific band is denoted by an asterisk. Blots are representative of at least four separate experiments.