Fig. 8. Validation of the SH-SY5Y stable cell line over-expressing α-Syn-EGFP and recovering of the DOPAL effect by treatment with Lys C-dots. (A) Immunofluorescence of SH-SY5Y stable cells overexpressing α-Syn-EGFP (green) and stained with an anti-α-Syn (MJFR1) antibody (red) to display co-localization. Nuclei are stained with Hoechst (blue), scale bar 50 μM. (B) Western blot of SH-SY5Y cells over-expressing α-Syn-EGFP treated with 100–200 μM DOPAL at different time points (18–24 hours). DOPAL induced dimers revealed by the anti-α-Syn (MJFR1) antibody are detected at 90–100 kDa. (C) Western blot analysis of lysates of SH-SY5Y cells over-expressing α-Syn-EGFP pre-treated with specified concentrations of Lys C-dots for 24 hours, following 150 μM DOPAL treatment for 18 hours. (D) In the quantification, the band intensity of the α-Syn-EGFP dimer was normalized to HSP70. Data are presented as mean ± SEM (n = 3).