FIGURE 1.

WTS binds directly to cytoskeleton proteins. A, scheme of the α-Syn in vitro aggregation protocol to generate α-Syn multimers (top). Multimers of WTS and different α-Syn variants were analyzed by electron microscopy (bottom). Scale bar, 200 nm. Nomenclature of different monomeric and multimeric α-Syn forms is shown (bottom). B, representative ligand blot performed by spotting 1 μg of the respective cytoskeleton proteins onto the nitrocellulose membrane strip. BSA was used as a negative control. 3 μl of 100 μm α-Syn solution served as a positive control (Pos. Control). Each strip was incubated in 100 μm α-Syn multimer solution and developed with anti-α-Syn antibody and fluorescently labeled secondary antibody. α-Syn proteins are shown on the left. PBS, the stripe that was incubated in PBS instead of α-Syn solution. C and D, densitometry analysis of three ligand blot experiments with α-Syn monomers (C) or multimers (D), respectively. A higher level of binding of WTS to cytoskeleton proteins as compared with different α-Syn variants was observed. Binding was stronger for α-Syn multimers than for monomers. Bars, mean intensity ± S.D. (error bars). *, p < 0.05.