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Probing Endocytosis During the Cell Cycle with Minimal Experimental Perturbation

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Book cover Clathrin-Mediated Endocytosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1847))

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.

The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-8719-1_19

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Change history

  • 24 October 2018

    This book was inadvertently published with the incorrect title as Clathrin-Mediated Endoytosis: Methods and Protocols. This has now been corrected throughout the book to Clathrin-Mediated Endocytosis: Methods and Protocols.

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Acknowledgments

A.J.M.S. was supported by Fundação para a Ciência e Tecnologia (SFRH/BD/33545/2008), and E.B. was a Biotechnology and Biological Sciences Research Council (BBSRC) David Phillips Research Fellow.

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Authors and Affiliations

  1. Cell and Developmental Biology Program, Centre for Genomic Regulation (CRG), Barcelona, Spain

    António J. M. Santos

  2. Institute of Structural and Molecular Biology, University College London and Birkbeck, London, UK

    Emmanuel Boucrot

Authors
  1. António J. M. Santos
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  2. Emmanuel Boucrot
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Corresponding author

Correspondence to Emmanuel Boucrot .

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Editors and Affiliations

  1. Department of Cellular & Molecular Physiology, University of Liverpool, Institute of Translational Medicine, Liverpool, UK

    Laura E. Swan

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Santos, A.J.M., Boucrot, E. (2018). Probing Endocytosis During the Cell Cycle with Minimal Experimental Perturbation. In: Swan, L. (eds) Clathrin-Mediated Endocytosis. Methods in Molecular Biology, vol 1847. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8719-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-8719-1_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8717-7

  • Online ISBN: 978-1-4939-8719-1

  • eBook Packages: Springer Protocols

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