Fig. 3.

High-throughput screening identifies three kinase inhibitors with potent preventative effects on α-synuclein seeding. a Schematic workflow of kinase inhibitor screening using NPR-H001 cells. Assay plates seeded with reporter cells are pre-treated with inhibitors prior to PFF addition and analyzed by flow cytometry after 48 h. b Resulting z-scores from kinase inhibitor screening distributes as a waterfall plot with the majority displaying little effect and few tail-end candidates showing large alterations. c Ranked representation of samples with z-scores beyond the 1.5 threshold identifies compounds with potential impact on α-synuclein aggregation. d Single compound validations show a visual reduction in formed α-syn aggregates 48 h following PFF addition (scale 40 µm) e, with differences already being detectable 20 h post PFF addition e. f Using NPR-H-001 reporter cells to confirm the significant reduction in induced aggregation (Enza: p < 0.001, GF: p < 0.001, SB80: p < 0.001, SB90: p < 0.001) following compound addition by flow cytometry. Bar chart shows mean ± SD, ***p < 0.001. Statistical testing was performed using one-way ANOVA with Dunnett’s T3 post hoc test for multiple comparisons to a control group