Figure 7. Inhibitors reduce the seeding by MSA derived α-syn fibrils.

(A) Experimental design of cell culture seeding assay showing MSA derived α-syn fibrils (green) were incubated with each inhibitor (red) for 3–4 hr to allow binding. Then HEK293 cells expressing YFP-labelled A53T α-syn were transfected with this mixture. After 7 days of transfection cells were imaged using fluorescence microscope and the bright fluorescent puncta were counted using imaging cytometer. (B and C) Representative fluorescence micrographs of the cells show that all the four inhibitors cause reduction in puncta (shown by white arrows) both in MSA1 (B) and MSA2 (C) transfected cells. Scale 100 µm. (D and E) Quantification of puncta in different conditions. All four inhibitors significantly reduce seeding by both MSA1 and MSA2 samples. Results shown as Mean + SD (n = 3) of technical replicates. Statistical significance was analyzed by two-way ANOVA. (**p<0.01,***p<0.001,****p<0.0001).