FIGURE 4.

Incorporation of Dan-Aβ42 monomer into replicated oligomers after seeding. A, schematic for incorporation of Dan-Aβ42 into replicated LFAOs after seeding for 72 h. B, SEC fractionation of 50 μm Dan-Aβ42 seeded with 1 μm LFAO (2% seed) for 72 h at room temperature. C, immunoblot showing the comparison of seeded sample before and after SEC. Lane 1, 50 μm Dan-Aβ42 monomer (1.8 μg); lane 2, LFAO seed (36 ng); lane 3, Dan-Aβ42 control in the absence of seeds after 72 h; lane 4, aliquot of the seeding reaction immediately after incubation at 0 h; lanes 5 and 6, the total reaction and supernatant of the reaction, respectively; lanes 7–9, SEC fractions 16, 17, and 18 show replicated oligomers (arrows); lane 10, monomer fractionated in the included volume (fraction 24). The LFAO seed amount of 36 ng was kept constant in the immunoblot. D, fractionated sample. Fractions 16, 17, and 18 show the Dan-Aβ42 emission at 450 nm upon exciting at 350 nm with both excitation and emission slits set 10 nm each. The results are one of three reproducible data sets. F, fluorescence; a.u., absorbance units.