Fig. 6: Differential processing of α-Syn polymorphs by the protein clearance pathways.

Phasor plots of the fluorescence lifetime of F65-Atto647 and F91-Atto647 seeds upon cells treatment with chloroquine (CQ) (a), MG132 (d), or DNAJB1 siRNA (g). Histograms of F65-Atto647 and F91-Atto647 showing the relative abundance (rel. abundance) of pixels with a certain fluorescence lifetime (ranging from 0 to 1.8 ns), averaged over all acquired images (n = 15 for each condition, mean ± SEM). b, e, h The mean fluorescence lifetime (τ) of F65-Atto647 and F91-Atto647 upon indicated treatment. Data are shown as mean ± SEM. Statistical analysis was done using unpaired t-test (for F65 b, h, F91 b, e, f) or Mann-Whitney test (for F65 e). Quantification of cells with visible foci (n = 30, mean ± SEM). Statistical analysis was done using a Mann-Whitney test (for F65 c, F91 c, f, i) or an unpaired t-test (for F65 f, i). Inhibition of autophagy (c) and the proteasome (f), and silencing of the HSP110/HSP70 disaggregation machinery by DNAJB1 siRNA (i) partially blocks the processing of F65-Atto647 seeds, which reduces α-Syn seeding. In contrast, only inhibition of autophagy partially blocks the processing of F91-Atto647 seeds.