Santos, AJM;
Boucrot, E;
(2018)
Probing Endocytosis During the Cell Cycle with Minimal Experimental Perturbation.
Methods in Molecular Biology
, 1847
pp. 23-35.
10.1007/978-1-4939-8719-1_3.
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Santos and Boucrot (Methods Mol Biol in press) Probing CME along the cell cycle - accepted.pdf - Accepted Version Download (1MB) | Preview |
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.
| Type: | Article |
|---|---|
| Title: | Probing Endocytosis During the Cell Cycle with Minimal Experimental Perturbation |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1007/978-1-4939-8719-1_3 |
| Publisher version: | https://doi.org/10.1007/978-1-4939-8719-1_3 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | 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 |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10057848 |
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