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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) Schematic representation of the CRISPR strategy used to introduce the 4E7E mutation. The green box indicates CyclinB1’s first exon, the blue box is a neomycin resistance cassette, grey box indicates the 5’UTR of CyclinB1. Yellow triangles indicate RoxP sites. The red text refers to CyclinB1’s ATG, blue text refers to PAM sites. (B) Graphs representing the pixel-by-pixel fluorescence intensity over a line going from centrosome to centrosome of RPE-1 Cyclin B1-mEmerald^+/+ compared to Cyclin B1^4E7E clones. For (panels B–G): n ≥ 12 cells per condition, N = 3 independent experiments. Mean ± standard deviation are plotted. (C, D) Cyclin B1 degradation graph representing the fluorescence intensity of Cyclin B1 over time of RPE-1 Cyclin B1-mEmerald^+/+ compared to Cyclin B1^4E7E clones. Mean ± standard deviation are plotted. (E) Dot plots representing the quantification of the mean fluorescence intensity of Cyclin B1 during metaphase in RPE-1 Cyclin B1-mEmerald^+/+ compared to Cyclin B1^4E7E clones. (F, G) Dot plots representing the maximal degradation speed (E) or the initial degradation time (F) of RPE-1 Cyclin B1-mEmerald^+/+ compared to Cyclin B1^4E7E clones. (H) Bar graph representing the karyotype classification from metaphase chromosome spreads of RPE-1 Cyclin B1-mEmerald^+/+ compared to p53^−/− clones and Cyclin B1^4E7E clones. The dotted line represents the level of aneuploidy in parental cells. N = 3 independent experiments. (I) Bar graph representing the karyotype classification from metaphase chromosome spreads of the indicated RPE-1 Cyclin B1^4E7E-mEmerald^+/+ clones following 3 weeks of ectopic expression of either Cyclin B1^WT-mScarlet or Cyclin B1^4E7E-mScarlet. N = 2 independent experiments. Source data are available online for this figure.