Parkinson’s Disease / 2011 / Article / Fig 4

Research Article

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

Figure 4

Characterization of alanine-76 in TCP1. (a) Live-cell GFP microscopy of TCP1 cells expressing WT, A76E, or A76R at 24 and 48 hours (left). Quantification: ~750 cells of each type were scored for these localization patterns: diffuse, aggregate, and aggregate/diffuse (right). Phenotypes were plotted as a percent of total cells that fluoresced ( ). (b) TCP1 cells expressing WT, A76E, or A76R were grown in inducing (EMM−T) media for 48 hours. Vector alone and GFP served as controls. Equal number of cells were serially diluted 5-fold and spotted onto repressing (EMM+T) or inducing (EMM−T) plates and grown for two days (right). The A76E and A76R mutants were equally toxic as WT ( ). (c) Western blot of TCP1 cells expressing WT, A76E, or A76R at 24 and 48 hours. Lysates were probed with anti-V5 (α-synuclein) or anti-β-actin (loading control). Expression is equivalent between WT and the alanine-76 mutants at 24 and 48 hours ( ).
392180.fig.004a
(a) Localization & Quantification
392180.fig.004b
(b) Toxicity
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(c) Expression

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