Fig. 8: Cytotoxicity in SH-SY5Y cells and primary neuron pathology model demonstrate hit compound rescue of αSN induce pathology.

a Overexpression of αSN in SH-SY5Y cells results in reduced cellular viability (~44%, red dashed line) as determined by CytoTox-Glo cytotoxicity assay. Treatment with control compound Anle138b as well as hit compounds RO and DEM demonstrate a dose-dependent rescue of toxicity with an EC₅₀ of 900, 78, and 65 nM for Anle138b, RO, and DEM, respectively. Biosensor hit BAY did not rescue the SH-SY5Y cell toxicity. b Using an αSN PFF-induced pathology model in primary mouse neurons, we evaluated the capacity of our hit compounds to rescue αSN PFF seed-associated pathology (phospho-S¹²⁹ staining). PFFs were treated at ~415 ng. Two of our hit compounds reduced the αSN pathology (BAY and DEM). Representative micrographs of each system are presented in Supplementary Fig. 14 with quantification of cytotoxicity via NeuN counting in Supplementary Fig. 15. N = 3 independent experiment; *p < 0.05, **p < 0.01, ns indicates not significant.