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Tracking the Cyclin B1-GFP Sensor to Profile the Pattern of Mitosis Versus Mitotic Bypass

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Cell Cycle Oscillators

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1342))

Abstract

This chapter provides a method for quantitative single cell analysis to track the transition of single cells from G2, indicated by high cyclin B1 levels, to G1 polyploidy phase (G1(p)), indicated by low cyclin B1 levels, in a 4n population. The cell tracking methodology described provides a fluorescence fingerprint suitable for deriving G2/M or G2/G1p transitions. Notably, during late G2 the absolute cyclin B1-eGFP reporter levels obtained were high and the switch-off point identifiable, with destruction rates of a similar order across all cell cycle routing avenues. The three principle parameters extracted were defined as (1) G2-to-G1(p) transition duration (tGFPoff); (2) rate of sensor destruction (kGFPoff), and (3) peak sensor expression (GFPpeak).

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Acknowledgement

This work was supported by the EPSRC grant EP/F040954/1 and the BBSRC Sparking Impact Award for LineageP.

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Authors and Affiliations

  1. Institute of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, Heath Park, Cardiff, CF14 4XN, UK

    Victoria Griesdoorn, Marie Wiltshire, Paul J. Smith & Rachel J. Errington

  2. College of Engineering, Swansea University, Singleton Park, Swansea, SA2 8PP, UK

    M. Rowan Brown

Authors
  1. Victoria Griesdoorn
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  2. M. Rowan Brown
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  3. Marie Wiltshire
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  4. Paul J. Smith
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  5. Rachel J. Errington
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Corresponding author

Correspondence to Rachel J. Errington .

Editor information

Editors and Affiliations

  1. Department of Oncology, University of Oxford, Oxford, United Kingdom

    Amanda S. Coutts

  2. Scientific writer, London, United Kingdom

    Louise Weston

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© 2016 Springer Science+Business Media New York

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Griesdoorn, V., Brown, M.R., Wiltshire, M., Smith, P.J., Errington, R.J. (2016). Tracking the Cyclin B1-GFP Sensor to Profile the Pattern of Mitosis Versus Mitotic Bypass. In: Coutts, A., Weston, L. (eds) Cell Cycle Oscillators. Methods in Molecular Biology, vol 1342. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2957-3_17

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  • DOI: https://doi.org/10.1007/978-1-4939-2957-3_17

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2956-6

  • Online ISBN: 978-1-4939-2957-3

  • eBook Packages: Springer Protocols

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