Fig. 2. CRBN interferes with the chaperonic activities of DJ1.

a Schematic representation of aggregation and disaggregation of α-SYN (adapted³²). b Turbidimetry assay of α-SYN aggregates (prepared from 30 mg/mL purified recombinant α-SYN protein) incubated with or without 15 μM of DJ2, DJ1, and CRBN as mentioned (with 30 μM Hsp70, 15 μM HSPA4 or (APG2) and 2 mM ATP added to all the samples) and monitored at 500 nm. c Disaggregation assay of α-SYN aggregates. ThT fluorescence of α-SYN fibrils (2 μM) in the presence of chaperones (1 μM DJ1, DJ2, or CRBN as mentioned with 2 μM Hsp70, 1 μM APG2 and 2 mM ATP added to all the samples, incubated at 30 °C for 2 h. d, e Aggregation assays for α-SYN monomers incubated with mentioned combinations with 2 μM Hsp70, 1 μM APG2, and 2 mM ATP added to all the samples, incubated at 30 °C for 150 h with gentle rotatory shaking. f Aggregation assays for α-SYN monomers as mentioned in (e), performed in the presence of different concentrations of DJ1 indicated in the legend. Student’s t test and one-way ANOVA was used for the statistical analysis of data. Experiments were repeated 3 times with 3 samples (n = 9) for all data points. Significance levels (p values) are shown in Supplementary Table 1.