Systematic Comparison of the Effects of Alpha-synuclein Mutations on Its Oligomerization and Aggregation
Figure 6
A. Native Gels. Immunoblot analysis of native PAGE of cells transfected with the BiFC constructs in HEK 293 cells. Smears indicate the presence of oligomeric species of ASYN with different sizes. n = 2. B. STED microscopy. Selected mutants were imaged in order to characterize the fine structure of the inclusions. C. Thioflavin S staining. H4 cells expressing selected SynT mutants were incubated with ThioS in order to reveal beta sheet-rich structures. Some of the inclusions display amyloid-like properties, with increased staining in the inner part of the inclusions, indicated with arrow heads (▸). Scale bar: 10 µm. D-E. Triton X-100 solubility assay and quantification. H4 cells show that all mutants form detergent insoluble species. Student's t test (*p<0.05, **p<0.01, ***p<0.001). n = 2. Quantification of insoluble fraction shows a decrease in TP and E57K mutants.