Chromothripsis pp 159-168 | Cite as

Experimental Determination of Checkpoint Adaptation by Mitotic Shake-Off and Microscopy

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1769)

Abstract

Cells that undergo checkpoint adaptation arrest at and then abrogate the G2/M cell cycle checkpoint to enter mitosis with damaged DNA. Cells surviving this process frequently contain micronuclei, which can lead to genomic change and chromothripsis. In this chapter we describe how to induce checkpoint adaptation and detect it by time-lapse video and immunofluorescence microscopy and how to isolate cells undergoing checkpoint adaptation from a total cell population.

Key words

Checkpoint adaptation Checkpoint kinase 1 Cyclin-dependent kinase 1 DNA damage HT-29 cells Micronuclei Microscopy Mitosis Mitotic shake-off 

Notes

Acknowledgments

Our research is funded by the Canada Foundation for Innovation (CFI) and the Alberta Innovates Sustainability Fund. We thank the members of the Cancer Cell Laboratory, the University of Lethbridge, and Dr. Aru Narendran (University of Calgary) for the support.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.POETIC Laboratory for Preclinical and Drug Discovery Studies, Departments of Pediatrics and OncologyAlberta Children’s Hospital Research Institute, Arnie Charbonneau Cancer Institute, University of CalgaryCalgaryCanada
  2. 2.Cancer Cell Laboratory, Department of Biological SciencesUniversity of LethbridgeLethbridgeCanada

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