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. 2019 Sep 4;10:3987. doi: 10.1038/s41467-019-11773-x

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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. 6 — Poly-SUMO2/3 prevents accumulation of CCAN proteins at centromeres. a U2OS cells were transfected either with a pool of four siRNAs against SENP6 (siSENP6) or a pool of four nontargeting siRNAs (NTP). Cells were fixed and stained with Hoechst to visualize DNA and CENP-T antibody 2 days post transfection. Panels show representative pictures of mitotic (left panel) and interphase cells (right panel). Scatter plots show quantifications of the average CENP-T foci intensities per cell (for mitotic cells) or per picture (for interphase cells). A two-sided t-test was performed. ****p < 0.0001. n (NTP mitotic cells) = 15; n (siSENP6 mitotic cells) = 16; n (NTP interphase cells) = 16; n (siSENP6 interphase cells) = 16. Dashed lines indicate areas of DNA. b U2OS cells stably expressing GFP-CENP-W were treated as in panel a. Cells were fixed and stained with Hoechst to visualize DNA and GFP antibody to enhance GFP signal. Left panels show representative pictures of mitotic (left panels) and interphase cells (right panels). Scatter plots show quantifications of the average CENP-W foci intensities per cell (for mitotic cells) or per picture (for interphase cells). A two-sided t-test was performed. ****p < 0.0001; ***p < 0.001. n (NTP mitotic cells) = 15; n (siSENP6 mitotic cells) = 15; n (NTP interphase cells) = 16; n (siSENP6 interphase cells) = 16. c U2OS cells were treated as in panel a, fixed and stained with Hoechst to visualize DNA and CENP-A antibody 2 days post transfection. Panels show representative pictures of mitotic (left panel) and interphase cells (right panel). Scatter plots show quantifications of the average intensities of CENP-A foci per cell (for mitotic cells) or per picture (for interphase cells). A two-sided t-test was performed. ****p < 0.0001. n (NTP mitotic cells) = 16, n (siSENP6 mitotic cells) = 16; n (NTP interphase cells) = 16, n (siSENP6 interphase cells) = 16. Dashed lines indicate areas of DNA. Scale bars = 5 µm (mitotic cells), 10 µm (interphase cells). All error bars shown represent standard deviation (SD). Source data are provided as a Source Data file