Fig. 1
αS seeding can be sensitively detected by FRET in αS biosensor line. A Representative FRET confocal microscopy images of αS biosensor line (αS CFP/YFP) treated with Lipofectamine (left, ctrl), 10nM αS monomer (center) and αS PFF (right) after 24 hours. Scale bar= 10μm. B Flow cytometry tracing of FRET signal from αS biosensors. FRET+ gate (CFP vs FRET) was drawn from empty Lipofectamine treated cells with no aggregation. C Quantitation of integrated FRET signal from FRET-flow cytometry is calculated as % FRET+ cell * Median (FRET+ Intensity). **** p < 0.0001, n.s., no significance by one-way ANOVA. Error bars are ±S.E.M. D Quantitation of cells treated with increasing concentrations of αS PFF for 24 hours. Cells are harvested after 24 hours and analyzed the same as 1C. Each dot represents an independent experiment