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. 2017 Oct 18;7:13444. doi: 10.1038/s41598-017-13870-7

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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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Western blot analysis of the Rs2b and the different mutants from cell nuclei purified from infiltrated N. benthamiana leaves. His-tagged 2b proteins were detected with penta-his antibody and silver staining was used to monitor the equivalence of protein loading. Mutants SPS/40-42/AAA, SPS/40-42/APA, and single mutants SPS/40-42/APS, SPS/40-42/SPA and SPS/40-42/SAS were detectable from purified nucleoli with penta-his antibody. The mutants 2b/40-42/DPS, 2b/40-42/SPD and 2b/40-42/DPD were not detectable in nuclei. DAPI-staining was used for visualisation of the purified nuclei (appearing blue) with fluorescence microscope using UV-light. The bar represents 10 µm.