Fig. 5

Analysis of K18 sucrose gradient ultracentrifugation fractions. Fractions obtained by sucrose gradient ultracentrifugation of aggregated, and non-aggregated K18 protein were analyzed by Western blot under native (a) condition. Detection performed with anti-myc primary antibody and anti-mouse HRPO-labelled secondary antibody. (Top: non-sonicated aggregated K18; middle: sonicated aggregated K18; bottom: monomeric K18). Fraction 1 has the highest density; Fraction 19 has the lowest density. Fraction P is a resuspension of the pellet in PBS. In panel (b), an MSD analysis of corresponding fractions using the aggregate specific PT76/PT76 assay is represented. Functional analysis of K18 sucrose gradient fractions can be observed in panel (c). Seeding capacity in the absence of transfection reagent was evaluated by percentage of FRET positive cells. A representative profile of at least three independent experiments is shown. K18 Tau fibrils are analyzed by electron microscopy (d-e). Non-sonicated (d) and sonicated K18 Tau (e) samples are used for this analysis. Images are captured at 43000x magnification