α-Syn triggers ROS in aged <i>elo</i> cells.

<p>(A) Wild-type cells containing empty plasmid (pAG426GAL) or expressing WT α-syn (pAG426GAL-WT) were cultured for 24 h in inducing media (+gal) at 30°C. After 24 h, cells were incubated with DHR 123 (5 µg/ml) for 1 h, and then visualized by differential interference contrast (DIC) and fluorescence microscopy (DHR). Cells were counted and the percentage of cells exhibiting red fluorescence was plotted. (B) Plot of the percentage of cells (WT, <i>elo1</i>Δ, <i>elo2</i>Δ, and <i>elo3</i>Δ) that exhibited red fluorescence due to ROS accumulation. Cultures were grown for 24 h in inducing media at 30°C and then stained with DHR. Each value was obtained from two independent experiments, where the total number of cells counted was 600. Error bars reflect s.e.m. (C) Reduced glutathione (GSH) protects aged cells from WT α-syn. The various cells expressing WT α-syn were grown in selective media containing sucrose to stationary phase, and then cells were washed and resuspended in galactose media with or without 10 mM GSH. After 3 days (1 day on aging curve) incubation, aliquots were removed from culture, diluted onto plates containing rich media, incubated for 2 days, and then colonies were counted. Each bar represents the number of colonies at the 3 days divided by the number of colonies at time zero. Experiments were conducted three times, and error bars represent ± s.e.m.