FIGURE 6.

α-Synuclein interference with the activation of autophagy. A, shown is spotting analysis of wild-type (W303) yeast cells co-transformed with high copy plasmids carrying different untagged α-synuclein versions and with a high copy plasmid carrying GFP-Atg8 under the control of a copper-inducible promoter. B, shown is live-cell microscopy of W303 cells during WT α-synuclein induction before shifting to SD(−N) starvation medium (autophagy off) and after (autophagy on). Scale bar = 1 μm. C, Western blots and quantifications present autophagy percents in cells expressing untagged α-synucleins during 8 h of induction in SC medium supplemented with galactose at the indicated time points. The values represent the average of four independent experiments. Significance of differences was tested using one-way ANOVA test (P* < 0.002; P**, P*** < 0.02; n = 4). D, Western blots and quantifications present autophagy percents of cells preinduced for 4 h to produce α-synuclein, then shifted to nitrogen starvation medium SD(−N) further supplemented with galactose. Probes were taken at 2 and 4 h after the shift. Previous to the shift, cells had been induced in galactose-containing SC medium for 4 h. The columns represent the average of 4 independent experiments. For control, W303 cells expressing the empty α-synuclein vector and the pCuGFP-Atg8 plasmid were used. Significance of differences was tested using one-way ANOVA test (P*, P** < 0.0002; n = 4).