Fig. 5: α-syn distribution at the IMS upon inhibition of complex I inhibitor or oxidative stress or inhibition of endosome-lysosome system acidification.

HeLa cells were co-transfected with wild type a and mutants b, c α-synS11 and IMS GFP_1–10, and treated with 10 μM rotenone for 12 h, 250 μM H₂O₂ for 12 h or 50 mM NH₄Cl for 2 h at 37 °C in a 5% CO₂ atmosphere, stained with an anti α-syn antibody and observed at a confocal microscope at 488 and 633 nm excitation wavelength. d–f CTCF was quantified as described in the ‘Materials and Methods’ section and reported as normalized values with respect to WT α-syn. (d, Student’s t-test: ***p < 0.001, **p < 0.01, *p < 0.05; One-way ANOVA p=0.0037. Dunnett’s multiple comparison test retrieved a statistically significant difference only for the Rot treated group, ***p < 0.001; f Student’s t-test: *p < 0.05; One-way ANOVA and Dunnett’s multiple comparison test retrieved no statistically significant differences between the groups). Quantification of CTCF has been done on at least 36 cells from at least three independent experiments for each condition. Scale bar is 20 μm