Parkinson’s Disease / 2011 / Article / Fig 5

Research Article

Contribution of Alanine-76 and Serine Phosphorylation in α-Synuclein Membrane Association and Aggregation in Yeasts

Figure 5

Characterization of alanine-76 in BY4741. (a) Live-cell GFP microscopy of BY4741 cells expressing WT, A76E, or A76R at 24 and 48 hours (left). Quantification: ~750 cells of each type were scored for these localization patterns: cytoplasmically diffuse, and plasma membrane, aggregate, plasma membrane/diffuse, plasma membrane/aggregate (right). Phenotypes were plotted as a percent of total cells that fluoresced ( ). (b) BY4741 cells expressing WT, A76E, or A76R were grown in inducing (galactose) media for 48 hours. Vector alone and GFP served as controls. Equal number of cells were serially diluted 5-fold and spotted onto repressing (glucose) or inducing (galactose) plates and grown for two days (right). No toxicity was apparent in budding yeast ( ). (c) Western blot of BY4741 cells expressing WT, A76E, or A76R at 24 and 48 hours. Lysates were probed with anti-V5 (α-synuclein) or anti-PGK. Expression is equivalent between WT and the alanine-76 mutants at 24 and 48 hours.
(a) Localization & Quantification
(b) Toxicity
(c) Expression

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