FIGURE 7.

Parkin binds α-synuclein under stress and results in α-synuclein-associated protein ubiquitination. A, HeLa cells transiently expressing GFP-parkin WT or A240R were treated with DMSO control (−) or 1 μm CCCP (+) for 18 h. Endogenous α-synuclein was immunoprecipitated (IP) and the samples were immunoblotted (IB), as indicated. Ubiquitinated species are marked by brackets. B, HeLa cells stably expressing YPF-parkin were transfected with either control or ubc13 siRNA. 48 h after transfection, cells were treated with DMSO control, CCCP (0.5 μm), or rotenone (50 nm) for 18 h. Endogenous immunoprecipitation and whole cell lysate input were analyzed by Western blotting with anti-α-synuclein and other antibodies, as indicated.