Fig. 5

Membrane binding is essential for the aSyn internalization and inclusion formation. a and b Immunoblot and quantification of the comparison of WT aSyn and two mutants (A30P and A11P/V70P) with different membrane-binding properties using a biotinylation assay. Dotted bars refer to the band corresponding to aSyn dimers (aSyn**), and clear bars refer to aSyn monomers (aSyn*). c ICC and d immunoblotting of non-treated (NT) cells, or cells treated with WT, A30P, or A11P/V70P aSyn for 24 h. Scale bar: 30 μm. e Quantification of the immunoblot in panel D. Dotted bars refer to the band corresponding to aSyn dimers (aSyn**), and clear bars refer to aSyn monomers (aSyn*). Statistical tests were performed using one-way ANOVA with repeated-measures for grouped analysis, followed by Tukey’s post-hoc tests. Data were expressed as mean ± SEM and a 0.5% general significance level was defined, with significance levels as follows: *: p < 0.05; **: p < 0.01; ***: p < 0.001. Statistical significance is indicated with the symbol “#” for the monomers, “+” for the dimers, and “*” for the combination between monomers and dimers