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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 EV3 — (A) Structural superposition of Scc1-SA2 (purple and cyan) bound to CENP-U (orange) and CTCF (purple-blue). F44, F46 of CENP-U and Y226, F228 of CTCF are shown in stick. (B) Fo-Fc omit electron-density Fourier map contoured at 2.0 σ. Residues of CENP-U are shown in orange, and SA2 and Scc1 are in green and blue, respectively. (C) HeLa cell lysates were subjected to pull-down with GST or GST-CENP-U (1–60) in the forms of WT, ADA, and FKF, followed by immunoblotting with antibodies for Scc1 and SA2, and CBB staining. (D) Lysates prepared from HEK-293T cells transiently expressing CENP-U-GFP in the forms of WT, ADA, and the indicated fragments were subjected to pull-down with GST or GST-Scc1-SA2, followed by immunoblotting with the antibody for GFP, and CBB staining. (E) U2OS-LacO cells transiently expressing the indicated proteins were stained with antibodies for the Flag-tag, Myc-tag, and DAPI. Example images are shown. (F) U2OS-LacO cells transiently expressing the indicated proteins were stained with the antibody for Plk1, and DAPI. Example images are shown. (G) Lysates prepared from HEK-293T cells transiently expressing SFB-CENP-Q and CENP-U-GFP (WT or ADA) were subjected to pull-down with GST or GST-Scc1-SA2, followed by immunoblotting with antibodies for GFP and the Flag-tag, and CBB staining. Data information: The white arrows point to the LacO repeats (E, F). Scale bars, 10 µm (E, F). Irrelevant lanes were removed (C, D, G).