Fig. 2

Dot blot immunoassay of αSyn_RT-QuIC products from OM-seeded reactions. Representative dot blot images of αSyn_RT-QuIC end products from OM samples of patients with PD (n = 2), MSA-P (n = 1), MSA-C (n = 1), and from HS (n = 1) obtained using an antibody against αSyn filament or antibody against total αSyn (a). The αSyn_RT-QuIC product of an autopsy skin sample from a PD case (PD-as) was used as a positive control [18]. The figure shows that αSyn filament immunoreactivity was present only in the products generated by OM samples of PD and MSA-P but was instead absent in those produced by MSA-C and HS. The total αSyn immunoreactivity was observed in all OM samples. Statistical analysis revealed significantly higher levels of αSyn filament in αSyn_RT-QuIC products generated by OM from PD (p < 0.01) and MSA-P (p < 0.0001) compared to those of MSA-C and HS (one-way ANOVA) (b). Total αSyn levels were comparable between groups (p = 0.0619, one-way ANOVA) (c). The signal intensity of each dot was normalized to that of HS controls (set to the unit of 1) and represented as mean with SD in (b) and (c). Densitometric analyses in (b) and (c) have been performed on dot blot images of αSyn_RT-QuIC products generated from PD (n = 13), MSA-P (n = 20), MSA-C (n = 10) and HS (n = 11) OM samples at USA-lab (see Supplementary Fig. 3b)