Figure 5
Self-assembly process for Sup35-NM.
(A) Time-course tracking of Sup35-NM self-assembly process as monitored by SEC. The Sup35-NM solution was analyzed at 2 h (blue), 4 h (magenta), 8 h (orange), 12 h (cyan), 20 h (red), 28 h (green), and 32 h (pink). (B) TEM images of 46 min (size determined from 50 spherical-shaped structures) and 41 min (size determined from 100 fibrils) species from SEC. (C) Western blots for Sup35-NM monomer (in 8 M urea, spot 1) and 41 min (spot 2), 46 min (spot 3), 73 min (spot 4) species from SEC. An anti-Sup35-NM, sequence-specific antibody was used for the NM lane. An oligomer-specific antibody was used for the A11 lane21,25,29. (D) Sup35-NM self-assembly process, as monitored by Congo red binding experiment, in the absence (blue) and presence of short fibril (red) or FP fibril (green).
