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. 2020 Nov 20;10:20295. doi: 10.1038/s41598-020-77164-1

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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N17 mutations in recipient cell lines altered sensitivity of seeding-induced mHTT aggregation. (a) A diagram illustrates the constructs of GFP-tagged mHTT-exon 1 (46Q) fragments, which are used to generate stable cell lines. (b) A diagram illustrates the generation of mHTT aggregates (fibrils) and seeds from monomeric recombinant mHTT-51Q proteins. (c) Cell lines generated by stably transfected HEK-293 cells with GFP tagged mHTT-exon 1 (46Q) variants were used as recipient cells of the seeding assay. Constructs of the transgenes are labeled on the left of the panel. Recombinant mHTT-exon 1 (51Q) in the forms of monomers, fibrils and seeds were added directly to the cultured cells for 3 days. Representative confocal images of the cells are shown. Bright green punctae are intracellular mHTT-GFP aggregates induced by seeding. N/T = non-treated. (d,e) The number of HTT-GFP aggregates was quantified using ImageJ and presented as mean ± s.e.m., n = 5–10, **p < 0.01.