Experimental Determination of Checkpoint Adaptation by Mitotic Shake-Off and Microscopy
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 wordsCheckpoint adaptation Checkpoint kinase 1 Cyclin-dependent kinase 1 DNA damage HT-29 cells Micronuclei Microscopy Mitosis Mitotic shake-off
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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