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. 2017 Jul 19;7:5853. doi: 10.1038/s41598-017-05829-5

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© The Author(s) 2017

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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Sup35-N5Y strongly rescues the [PSI ⁺] induction defect. (A) [PSI ⁺] induction experiments demonstrate that three Sup35 mutants restore [PSI ⁺] formation to nearly WT levels in rnq1-Q298R cells. These mutations do not increase [PSI ⁺] formation in a RNQ1 genetic background. Horizontal axis labels denote SUP35 genetic status. The [rnq ⁻] control cells express RNQ1 and SUP35. More than 13,000 colonies were assessed over five biological replicates. Error bars represent mean ± s.e.m. The “*” symbols represent a significant difference between [PSI ⁺] induction with WT Sup35 in rnq1-Q298R versus induction with the indicated sup35 mutants, p < 0.005. (B) Boiled gel assays allow separate migration of monomeric and aggregated materials, confirming that Sup35 aggregation is reduced in the SUP35 rnq1-Q298R background relative to SUP35 RNQ1 cells. Sup35 aggregation is restored in sup35-N5Y rnq1-Q298R cells. We attribute the slightly aberrant monomer bands as an artifact of the experimental procedure, as samples were unboiled (and likely non-denatured) as per the protocol, and Sup35 may shed unevenly from aggregates in addition to the soluble pool. Western blot is a representative image from three independent experiments.