Journal: bioRxiv

Article Title: A new therapeutic approach for Parkinson’s disease: dual targeting of α-Synuclein aggregation and microglial function by the novel immunomodulator 3-Monothiopomalidomide

doi: 10.64898/2026.03.26.714051

Figure Lengend Snippet: ( A ) ThT fluorescence intensity monitored during α-Syn (100 μM) aggregation at 37 °C, in PBS at pH 7.4 and under orbital shaking (200 rpm). Kinetic traces for isolated α-Syn (black) and in co-incubation with POM (yellow) and 3MP (orange) are reported. All conditions were tested in triplicate. The intrinsic fluorescence of 3MP was found to be negligible compared to the intensity of ThT fluorescence. ( B ) t½ values of the kinetic curves. ( C ) Plateau values of the kinetic curves. One-way ANOVA followed by Dunnett’s post-hoc test was performed against protein alone (control). ns = not significant, * p < 0.05, *** p < 0.001. Shapiro–Wilk and Bartlett’s tests confirmed normality and homogeneity of variance. ( D ) Transmission electron microscopy (TEM) images of samples post 120h of incubations. The images were taken at both 100 nm and 200 nm magnifications. Samples were prepared using negative staining with 2% phosphotungstic acid (PTA) on carbon-coated 200 mesh copper grids. ( E ) Representative images of ThS staining on SH-SY5Y cells challenged with α-Syn pre-formed fibrils and treated with POM and 3MP (magnification 63x; scale bar: 20µm). ( F ) Quantification of ThS + signal expressed as ThS□ volume per cell (µm³/cell) in SH-SY5Y cells challenged with α-Syn pre-formed fibrils and treated with POM or 3MP. Data are shown as mean ± SEM from three independent biological replicates. One-way ANOVA followed Tuckey’s post-hoc test.

Article Snippet: To monitor the aggregation kinetics via ThT fluorescence, we employed a FLUOStar Omega (BMG Labtech) plate reader with excitation and emission wavelengths set at 448 nm and 482 nm, respectively.

Techniques: Fluorescence, Isolation, Incubation, Control, Transmission Assay, Electron Microscopy, Negative Staining, Staining