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. 2019 Jun 25;9:9210. doi: 10.1038/s41598-019-45545-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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CRES exhibits nucleation-dependent amyloid assembly. (a) Thioflavin T fluorescence in 8.7 μM CRES monomer alone, CRES seed, and monomer and seed combined. Monomer to seed molar ratio was 1:1.4. Reactions were done in 20 μM Thioflavin T, 11 mM MES, 20 mM HEPES, 112.5 mM NaCl, and 0.45 mM EDTA, pH 7.4. Fluorescence was measured continuously for 1 hour at 25 °C without shaking and then every hour for 5 hours. Because some evaporation occurred when plates were read overnight, after 5 hours plates were stored covered at RT and read the following day until 24 hours total time. The data show the mean ± SEM from n = 3 experiments using 3 independent CRES preparations. The error bars for the monomer are too small to be detected. (b) Negative stain TEM of samples in (a) including monomer and seed alone before being added to the ThT reaction (start) and after 24 hours in the ThT reaction (24 hr), and seeded reactions after 24 hours. Monomer, green; seed, red; monomer and seed combined, blue.