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. Author manuscript; available in PMC: 2017 Jul 4.
Published in final edited form as: Mol Psychiatry. 2017 Jan 3;22(9):1327–1334. doi: 10.1038/mp.2016.230

Figure 1. In vitro Aβ aggregation is enhanced by IAPP heterologous seeding.

Figure 1

A: Soluble seed-free Aβ40 (2000 nM) in 100 mM Tris-HCl pH 7.4 (200 μl) containing 5 μM thioflavin T (ThT) was allowed to aggregate at 22°C with intermittent shaking at 500 rpm in the absence or in the presence of either 20 nM Aβ40 seeds (homologous seeding) or 20 nM IAPP seeds (heterologous seeding). The extent of aggregation was periodically monitored by measuring the ThT fluorescence at excitation of 435 nm and emission of 485 nm. Error bars indicate standard deviation (S.D.) B: Transmission electron microscopy microphotography of double immuno-gold labeling of fibrils formed by the addition of IAPP seeds (10% with respect to Aβ) into seed-free Aβ1–42 (37 μM) aggregation solution. After in vitro cross-seeding, fibrils were stained using antibodies coupled to gold nanoparticles. IAPP was detected with an antibody coupled to 12 nm gold particles while Aβ was labeled with 6 nm gold particles. Scale bar: 100 nm. Red arrows point to aggregates showing staining for both peptides.