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. 2024 Apr 10;43(11):2166–2197. doi: 10.1038/s44318-024-00093-6

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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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(A) Western blot of whole-cell extracts prepared from control or DNAJC9-depleted cells with or without MCM2 depletion showing the efficiency of DNAJC9 and MCM2 depletions in HeLa YFP-CENP-A^Low. Alpha-tubulin was used as the loading control. Representative images from three biological replicates are shown. (B) MCM2 depletion suppresses CENP-A mislocalization in DNAJC9-depleted HeLa YFP-CENP-A^Low cells. Representative images of metaphase chromosome spreads immunostained for CENP-A in control or DNAJC9.3 siRNA-transfected cells with or without MCM2 depletion in HeLa YFP-CENP-A^Low from three biological replicates. Scale bar: 15 µm. Scale bar for inset: 5 µm. (C) Scatter plots showing CENP-A intensities corrected for background at the centromeric or noncentromeric regions as described in B. Each dot represents value from the individual chromosome. Chrs represent the total number of chromosomes measured per condition from three biological replicates. Red horizontal lines represent mean signal intensity and error bars represent SD across all measurements from three biological replicates. P values were derived from one-way ANOVA with Tukey’s ad hoc test. Source data are available online for this figure.