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. 2019 Jan 21;10:359. doi: 10.1038/s41467-018-08180-z

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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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Fig. 2 — LEF-10 functionally replaces the prion domain of the yeast Sup35 protein. a Yeast ade1-14 cells expressing LEF-10-Sup35MC were spread on complete (1/4 YPD) medium. [LEF⁺] strains formed white colonies which were distinguishable from [lef⁻] strains by the red pigment accumulated through the adenine biosynthetic pathway (top). Both phenotypes could be sustained during cell propagation (middle). [lef⁻] strains spontaneously generated [LEF⁺] colonies at a very low frequency (bottom, a colony with the [LEF⁺] phenotype is indicated by an arrow) and these [LEF⁺] colonies displayed Ade⁺ (suppression of ade1-14) phenotype when tested directly on SD-Ade medium. b Comparable characteristics of LEF-10-Sup35MC and full-length Sup35 in yeast. Serial dilutions of [LEF⁺] and [lef⁻] strains were spotted on the 1/4 YPD plate (the top panel), or medium lacking adenine (SD-Ade) on which only prion-containing i.e., [PRION⁺] strains suppressing the stop codon in the yeast ade1-14 allele could grow (the second panel). SDS-resistant aggregates in cell lysates of yeast strains expressing LEF-10-Sup35MC were examined by SDD-AGE (the third panel). The expression levels of full-length Sup35 and LEF-10-Sup35MC were examined by Western blot, probing with a Sup35C-specific antibody (the fourth panel). Endogenous phosphoglycerate kinase 1 (PGK1) was detected with a PGK1-specific antibody and served as a loading control (the bottom panel). [PSI⁺] and [psi^-] strains were used as positive and negative controls