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    Probing Endocytosis during the cell cycle with minimal experimental perturbation

    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.

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    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: School of Science > Biological Sciences
    SWORD Depositor: Mr Joe Tenant
    Depositing User: Mr Joe Tenant
    Date Deposited: 18 Sep 2018 10:34
    Last Modified: 28 Jul 2020 09:43
    URI: https://eprints.bbk.ac.uk/id/eprint/23826

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