Skip to main content
. Author manuscript; available in PMC: 2022 May 14.
Published in final edited form as: J Mol Biol. 2021 Mar 24;433(10):166953. doi: 10.1016/j.jmb.2021.166953

Figure 4.

Figure 4.

Cross-seeding of oligomers and sonicated fibrils with monomers. a) DLS analysis of DOPAL-derived αS oligomers isolated from SEC. (inset) SEC fraction containing the oligomer ‘o’ used in the study; ‘m’ refers to control monomer. b) CD spectra and AFM height image (inset) of DOPAL-derived αS oligomers used for cross-seeding reaction (scale bar = 1μm). Cross-seeding reactions with monomers of 15 μM TDP-43 PrLD alone (▲) or in the presence of 0.5 μM (■) or 1 μM (●) DOPAL-derived αS oligomers in the presence of 10 μM ThT. (inset) immunoblot of the reaction after 12 h probed with TDP-43 antibody; ‘c’ refers to TDP-43 PrLD monomer control. d) Seeding of 20 μM αS monomers with 4 μM (○), 2 μM (◇), and 1 μM (△) of αS sonicated fibrils, and 20 μM TDP-43 PrLD monomers seeded with 4 μM (●), 2 μM (♦), 1 μM (▲) of αS sonicated fibrils. e) Seeding of 20 μM TDP-43 PrLD monomers with 4 μM (○), 2 μM (◇), and 1 μM (△) of TDP-43 PrLD sonicated fibrils and seeding of 20 μM αS monomers with 4 μM (●), 2 μM (♦), 1 μM (▲) of TDP-43 PrLD seed. f) FTIR analysis of cross-seeding reactions from (c-e). TDP-43 PrLD monomers cross-seeded with 1 μM DOPAL-derived αS oligomers after 12 h of incubation (—); TDP-43 PrLD monomers seeded with 1 μM αS sonicated fibrils (—); αS monomers seeded with 1 μM TDP-43 PrLD sonicated fibrils (—) along with controls such as αS monomers (—) and TDP-43 PrLD monomers (—) (g-j) AFM height image of αS sonicated fibrils (g), αS sonicated fibrils seeded TDP-43 PrLD monomers (h), TDP-43 PrLD sonicated fibrils (i), and TDP-43 PrLD sonicated fibrils seeded αS monomers (j) (scale bar = 1 μm, inset = 200 nm).