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. 2024 May 7;43(12):2424–2452. doi: 10.1038/s44318-024-00104-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Figure EV4 — (A) Asynchronous HeLa cells stably expressing siRNA-resistant CENP-U-GFP (WT or the ADA mutant) were subjected to immunoblotting with antibodies for GFP, CENP-U, and α-Tubulin. (B) The indicated stable cell lines were immunostained with antibodies for GFP, CENP-C, and DAPI. Example images are shown. (C, D) HeLa cells stably expressing the indicated proteins were transfected with control siRNA or CENP-U siRNA. At 48 h post-transfection, cells were treated with MG132 for 8 h, then mitotic chromosome spreads were stained with the CENP-C antibody and DAPI. The percentage of cells in which the majority of sister chromatids was separated or unseparated was determined in 300 cells for each condition from three independent experiments, with statistics being analyzed for cells with separated chromatids using unpaired Student’s t-test. Means and SDs are shown (C). Example images are shown (D). Data information: Scale bars, 10 µm (B, D).