Figure 7

Rotenone induced α-syn oligomers and fibrils alter mitochondrial membrane potential and induce apoptosis. The cell viability on SH-SY5Y cells after 24 h treatment with α-syn aggregated species (O,OR,F,FR) at different concentrations(1–10 µM), rotenone (ROT, 5 µM) and SDS (0.2%) were assessed with MTT reduction assay (a) Percent cell viability is calculated to vehicle cell population normalized to 100%. Effect of α-syn aggregated species (O,OR,F,FR, 1 µM), rotenone (ROT, 5 µM) and valinomycin (0.3%) on mitochondrial membrane potential (ΔΨm) in SH-SY5Y cells measured by JC-1 staining (b). After 24 h treatment the cells were analyzed by flow cytometry and the quantitative ratio of red to green fluorescence is calculated. Late apoptosis induced by α-syn oligomers (O,OR, 1 µM) and fibrils (F,FR, 1 µM) measured by propidium iodide (PI) uptake (c). Flow cytometry analysis of apoptosis induced by the above mentioned treatments for48h after double labeling with PI and Annexin-V. Statistical significance was calculated by two tailed t-test and One way ANOVA analysis data expressed as means and ± SE (n = 3), *P < 0.05; **P < 0.01; ***P < 0.001;****P < 0.0001. (Supplementary information: Fig. S5a,b).