Fig. 1

Characterization of recombinant 2N4R Tau aggregates. A Fibril formation kinetics of recombinant human 2N4R Tau by Thioflavin T (ThT) fluorescence, fitted with the Finke-Watzky model of two-step nucleation-autocatalysis. B Dot-blot analysis of Tau aggregates at different time points during the fibrillization process in A by three different conformation-sensitive Tau antibodies (MC1, TNT-1 and TOMA). C Atomic-force microscopic image of a mixture of aggregates after 72 h of fibrillization, including large fibrils (filled arrowheads), small fibrils (open arrowheads), and oligomers (arrows). Scale bar: 200 nm. D Size-exclusion chromatography (SEC) analysis of the soluble fraction of aggregates after removing insoluble fibrils by ultracentrifugation, showing the absorbance at 214 nm in the eluting fractions, including Tau oligomers (Oligo) and monomers (Mono). E Dot-blot analysis of SEC fractions in D using the antibodies Tau5 (total Tau) and MC1 (conformationally altered Tau). F Dynamic light scattering measurements of Mono and Oligo Tau showing the hydrodynamic size distribution of soluble Tau species obtained from SEC (d.nm: diameter in nanometers)