Fig. 3
VCP inhibition enhances αS seeding. A αS biosensors were treated with scrambled control siRNA or VCP siRNA for 48 hours prior the application of αS monomer or PFF (10nM) with or without Lipofectamineand measured for FRET efficiency at 24 and/or 48 hours. N=4 for each groups.; n.s.= no significant, **** p < 0.0001 by two-way ANOVA compared with scramble siRNA in each conditions. error bars are ±S.E.M.). B αS biosensors were treated with Dynogo4a for one hour prior to αS PFFormonomer (10nM) treatment with Lipofectamine. n =12 repeats for PFF treated conditions. **** p < 0.0001; n.s. for monomer treated pairs by two-way ANOVA. C αS biosensors treated with αS PFF for 4 hours followed by media exchange and washout. FRET signal was measured the same as in 3A, 20 hours after washout. n.s. no significant and **** p < 0.0001 compared to vehicle washout at different concentrations. Each dot represents an independent experiment. D Normalized FRET of αS biosensors co-treated with αS monomer or PFF (30nM) with the indicated chemical compound or vehicle for four hours followed by washout and FRET measurement 20 hours later. (n =14 repeats for each PFF treated conditions; n.s. for all monomer treated pairs; **** p < 0.0001 by two-way ANOVA in some PFF treated pairs as indicated. error bars are ±S.E.M.). E Normalized FRET of αS biosensors co-treated with αS monomer or PFF (10nM) with the indicated chemical compound or vehicle for 24hours. FRET signal was obtained same as3A (n ≥8 repeats for each PFF treated conditions; n.s. for all monomer treated pairs; **** p < 0.0001 and * p < 0.05 by two-way ANOVA in some PFF treated pairs as indicated. error bars are ±S.E.M.)