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

Lucy H Swift; Roy M Golsteyn

doi:10.1007/978-1-4939-7780-2_10

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

Methods Mol Biol. 2018:1769:159-168. doi: 10.1007/978-1-4939-7780-2_10.

Authors

Lucy H Swift¹, Roy M Golsteyn²

Affiliations

¹ POETIC Laboratory for Preclinical and Drug Discovery Studies, Departments of Pediatrics and Oncology, Alberta Children's Hospital Research Institute, Arnie Charbonneau Cancer Institute, University of Calgary, Calgary, AB, Canada.
² Cancer Cell Laboratory, Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada. roy.golsteyn@uleth.ca.

PMID: 29564823
DOI: 10.1007/978-1-4939-7780-2_10

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.

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptation, Biological*
Cell Cycle Checkpoints*
Cell Line
Humans
Microscopy*
Microscopy, Fluorescence
Mitosis*
Time-Lapse Imaging