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. 2018 Aug 3;9:3057. doi: 10.1038/s41467-018-05354-7

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© The Author(s) 2018

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. 5 — HeV M protein expression does not impact the structure or distribution of Treacle-enriched compartments. HeLa cells expressing GFP alone (a) or the indicated proteins (b) were fixed and immunolabeled for Treacle before analysis. a dSTORM single molecule localization microscopy (SMLM) was used to generate 3D images of Treacle-enriched subnucleolar compartments. Scale bar indicates 1 μm. b Fluorescence microscopy was used to identify cells expressing GFP (green) and immunostained for Treacle (red/yellow in merged image), which were then analyzed by SMLM to detect Treacle. Scale bars correspond to the indicated sizes. c–e Images such as those shown in Fig. 5b were used to determine (c) the area of Treacle-enriched compartments (mean ± s.e.m., n ≥ 427 compartments), (d) the number of nucleoli per cell (mean ± s.e.m., n ≥ 14 cells), (e) the number of Treacle-enriched compartments per nucleolus (mean ± s.e.m., n ≥ 57 nucleoli). Statistical analysis used Student’s t-test; NS not significant