Qualitative and quantitative analysis of endocytic recycling

James B. Reineke, Shuwei Xie, Naava Naslavsky, Steve Caplan

Research output: Chapter in Book/Report/Conference proceedingConference contribution

6 Scopus citations

Abstract

Endocytosis, which encompasses the internalization and sorting of plasma membrane (PM) lipids and proteins to distinct membrane-bound intracellular compartments, is a highly regulated and fundamental cellular process by which eukaryotic cells dynamically regulate their PM composition. Indeed, endocytosis is implicated in crucial cellular processes that include proliferation, migration, and cell division as well as maintenance of tissue homeostasis such as apical-basal polarity. Once PM constituents have been taken up into the cell, either via clathrin-dependent endocytosis (CDE) or clathrin-independent endocytosis (CIE), they typically have two fates: degradation through the late-endosomal/lysosomal pathway or returning to the PM via endocytic recycling pathways. In this review, we will detail experimental procedures that allow for both qualitative and quantitative assessment of endocytic recycling of transmembrane proteins internalized by CDE and CIE, using the HeLa cervical cancer cell line as a model system.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
Subtitle of host publicationSorting and Recycling Endosomes, 2015
EditorsWei Guo
PublisherAcademic Press Inc.
Pages139-155
Number of pages17
ISBN (Print)9780128028292
DOIs
StatePublished - 2015

Publication series

NameMethods in Cell Biology
Volume130
ISSN (Print)0091-679X

Keywords

  • Clathrin-dependent internalization
  • Clathrin-independent internalization
  • EHD1
  • Endocytic recycling
  • Endosome
  • Flow cytometry
  • MICAL-L1
  • Quantitative analysis
  • Transferrin

ASJC Scopus subject areas

  • Cell Biology

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  • Cite this

    Reineke, J. B., Xie, S., Naslavsky, N., & Caplan, S. (2015). Qualitative and quantitative analysis of endocytic recycling. In W. Guo (Ed.), Methods in Cell Biology: Sorting and Recycling Endosomes, 2015 (pp. 139-155). (Methods in Cell Biology; Vol. 130). Academic Press Inc.. https://doi.org/10.1016/bs.mcb.2015.04.002