Lopacr1280 library

Fluorescence dye labeled α-Syn preformed fibrils were generated as previously described^{19 (link)}. siRNA-mediated gene silencing was performed by lipofectamine RNAiMAX (Invitrogen) mediated transfection following the instruction from the manufacture. HEK293T cells were seeded in white 1536-well microplates that have transparent bottom (Greiner BioOne) at 2000 cells/well in 2 µL media and incubated at 37 °C overnight (~16 h). pHrodo red-labeled α-Syn fibrils were added at 400 nM final concentration to each well by a dispenser. After 1 h incubation, compounds from the LOPAC^R1280 library (Sigma) were titrated 1:3 in DMSO and dispensed via pintool at 23 nL/well to the assay plates. After 24 h of incubation, the fluorescence intensity of pHrodo red was measured by a CLARIOstar Plus plate reader (BMG Labtech). Data were normalized using cells containing 400 nM pHrodo red-labeled Syn fibrils as 100% and medium containing 400 nM preformed fibrils-pHrodo red as 0%.

Fluorescence dye labeled α-Syn preformed fibrils were generated as previously described^{19 (link)}. siRNA-mediated gene silencing was performed by lipofectamine RNAiMAX (Invitrogen) mediated transfection following the instruction from the manufacture. HEK293T cells were seeded in white 1,536-well microplates that have transparent bottom (Greiner BioOne) at 2,000 cells/well in 2 μL media and incubated at 37 °C overnight (~16 h). pHrodo red-labeled α-Syn fibrils were added at 400 nM final concentration to each well by a dispenser. After 1 h incubation, compounds from the LOPAC^R1280 library (Sigma) were titrated 1:3 in DMSO and dispensed via pintool at 23 nl/well to the assay plates. After 24 hours of incubation, the fluorescence intensity of pHrodo red was measured by a CLARIOstar Plus plate reader (BMG Labtech). Data were normalized using cells containing 400 nM pHrodo red-labeled Syn fibrils as 100% and medium containing 400 nM preformed fibrils-pHrodo red as 0%.