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. 2024 Aug 14;43(19):4324–4355. doi: 10.1038/s44318-024-00194-2

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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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(A) Representative maximum projections of fluorescence confocal images over time of an RPE-1 Cyclin B1-mEmerald^+/+ cell progressing through mitosis. Time is expressed as mm:ss. The scale bar corresponds to 10 μm. (B–D) Quantification of Cyclin B1 fluorescence levels (normalised raw integrated density, RID) over time in RPE-1 Cyclin B1-mEmerald^+/+ cells: n ≥ 18 cells per condition, N = 3 independent experiments. Mean ± standard deviation are plotted. (E) Representative fluorescence confocal images of RPE-1 Cyclin B1-mEmerald^+/+; APC8-mScarlet^+/+ cells. The scale bar corresponds to 10 μm. (F, G) Representative graph of the autocorrelation of Cyclin B1-mEmerald and APC8-mScarlet and the cross-correlation between the two at the chromatin (F) and in the cytoplasm (G). In this and all following FCCS graphs, dots = data, lines = fit; Cyan = Cyclin B1-mEmerald, magenta = APC8-mScarlet, yellow = cross-correlation between cyclin B1-mEmerald and APC8-mScarlet. (H–J) Dot plots representing the K_D between endogenous Cyclin B1-mEmerald and APC8-mScarlet in the indicated conditions: n ≥ 21 cells per condition, N = 3 independent experiments. In this and all following K_D dot plots, each dot corresponds to a single measurement, horizontal black lines represent median values. In this and all following figures, the orange dotted line indicates anaphase (panel B), the dotted horizontal line indicates 0 (panels F, G), CycB1 Cyclin B1, mEm mEmerald, mSc mScarlet, see Appendix Table S2 for statistical tests and p values. ns not significant, p > 0.05; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 for all figures. Source data are available online for this figure.