Imaging “Hot-Wired” clathrin-mediated endocytosis

Laura A. Wood, Stephen J. Royle

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Clathrin-mediated endocytosis (CME) occurs continuously at the plasma membrane of eukaryotic cells. However, when a vesicle forms and what cargo it contains are unpredictable. We recently developed a system to trigger CME on-demand. This means that we can control when endocytosis is triggered and the design means that the cargo that is internalized is predetermined. The method is called hot-wired CME because several steps and proteins are bypassed in our system. In this chapter, we describe in detail how to use the hot-wiring system to trigger endocytosis in human cell lines and how to image the vesicles that form using microscopy and finally, how to analyze those images.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Place of PublicationNew York, U. S. A.
PublisherHumana Press
Pages83-94
Number of pages12
ISBN (Print)9781493987177
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1847

Keywords

  • Chemically induced dimerization
  • Clathrin
  • Endocytosis
  • Image analysis
  • Immunofluorescence
  • Live cell imaging

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Wood, L. A., & Royle, S. J. (2018). Imaging “Hot-Wired” clathrin-mediated endocytosis. In Methods in Molecular Biology (pp. 83-94). (Methods in Molecular Biology; Vol. 1847). Humana Press. https://doi.org/10.1007/978-1-4939-8719-1_7