Figure 1.

Clearance of α-synuclein oligomeric intermediates. COS-7 cells expressing α-syn were treated with 100 nm rotenone for the indicated times and then incubated in fresh medium without rotenone for 1 or 2 d. A, Cells were extracted with PBS/1% Triton X-100, and the Triton-insoluble fractions were analyzed by Western blotting. The bottom panel shows densitometric analysis of the Western data. The relative density is obtained by calculating the percentage of remaining aggregates after the rotenone washout. B, Oligomeric intermediates (S) and mature fibrillar inclusion bodies (P) were separated before (-) and 1 d after (WO) the rotenone washout and were analyzed by Western blotting. C, Quantitation of the number of mature inclusion bodies. Before and after the rotenone washout the cells were fixed and labeled for α-syn by immunofluorescence. Cells with mature juxtanuclear inclusion bodies were counted in six different randomly selected areas, and the number of these cells was divided by the number of nuclei to obtain the percentage.