Fig. 3. DJ1 decreases the toxicity of PFF in CRBN-dependent manner.
a Western blot analysis of migration patterns of α-SYN monomers (1st lane) and PFFs generated without DJ1 (2nd lane), with DJ1 (3rd lane) and with DJ1 and CRBN (4th lane). b Differentiated SH-SY5Y cells were transfected with β-gal using ChariotTM reagent for protein transfection as positive control to confirm the transfection efficiency. c Differentiated SH-SY5Y cells were treated with monomeric (named as non-treated NT) or PFFs prepared with or without DJ2, DJ1, and CRBN as mentioned in the legend of Fig. 2. d For the samples mentioned in (c), MTT assay was performed after 48 h to assess cell viability. e Differentiated SH-SY5Y cells were transfected with Myc Synuclein and re-transfected with PFFs prepared with or without DJ2, DJ1, and CRBN using ChariotTM. Cells were immunostained with pSer129 antibody (pSYN) and LAMP (lysosomal marker) and analyzed with confocal microscope after 48 h. Quantification is shown in the (f) panel. Error bars represent the SEM. Student’s t test was used for quantification of data with a 95% significance level in Excel. n = 5, *p < 0.05, **p < 0.01, and ***p < 0.001.