Santos, A.J.M. and Boucrot, Emmanuel (2018) Probing Endocytosis during the cell cycle with minimal experimental perturbation. In: Swan, L.E. (ed.) Clathrin-Mediated Endoytosis: Methods and Protocols. Methods in Molecular Biology 1847. Springer, pp. 23-35. ISBN 9781493987177.
Abstract
Endocytosis mediates the cellular uptake of nutrients, modulates signaling by regulating levels of cell surface receptors, and is usurped by pathogens during infection. Endocytosis activity is known to vary during the cell cycle, in particular during mitosis. Importantly, different experimental conditions can lead to opposite results and conclusions, thereby emphasizing the need for a careful design of protocols. For example, experiments using serum-starvation, ice-cold steps or using mitotic arrest produced by chemicals widely used to synchronize cells (nocodazole, RO-3306, or S-trityl-L-cysteine) induce a blockage of clathrin-mediated endocytosis during mitosis not observed in unperturbed, dividing cells. In addition, perturbations produced by mRNA interference or dominant-negative mutant overexpression affect endocytosis long before cells are being assayed. Here, we describe simple experimental procedures to assay endocytosis along the cell cycle with minimal perturbations.
Metadata
| Item Type: | Book Section |
|---|---|
| Additional Information: | Series Print ISSN: 1064-3745 |
| Keyword(s) / Subject(s): | Cell cycle, Cell division, Cell synchronization, Clathrin-mediated endocytosis, Dynasore, Endocytosis, Knock-sideways, Mitosis, Nocodazole, Pitstop, Preincubation on ice, RO-0336, S-Trityl-L-cysteine, Serum starvation, siRNA |
| School: | Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences |
| SWORD Depositor: | Mr Joe Tenant |
| Depositing User: | Mr Joe Tenant |
| Date Deposited: | 18 Sep 2018 10:34 |
| Last Modified: | 02 Aug 2023 17:44 |
| URI: | https://eprints.bbk.ac.uk/id/eprint/23826 |
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