Single Cell Fluorescence Ratio Image Analysis for Studying ESCRT Function in Receptor Trafficking

Jalal M. Kazan, Gergely L. Lukacs, Pirjo M. Apaja, Arnim Pause

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

The endosomal sorting complexes required for transport (ESCRT) comprise a major trafficking pathway for plasma membrane proteins and are fundamental for ubiquitin-dependent cargo endocytosis. Here, we describe a method for studying the effect of ESCRT complexes on endo-lysosomal membrane trafficking and their role in receptor integrin α5β1 downregulation. Single cell fluorescence ratio image analysis (FRIA), using appropriate fluorescence probes, enables the measurement of dynamics of integrin α5β1 containing vesicles and represents a live cell-based method for studying the role of ESCRTs.

Original languageEnglish
Title of host publicationThe ESCRT Complexes
Subtitle of host publicationMethods and Protocols
EditorsEmmanuel Culetto, Renaud Legouis
Place of PublicationNew York, United States
PublisherHumana Press Inc.
Pages93-103
Number of pages11
ISBN (Electronic)9781493994922
ISBN (Print)9781493994915
DOIs
Publication statusPublished - 1 Jan 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1998
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • ESCRT
  • FRIA
  • HD-PTP
  • Integrin α5β1
  • Lysosomes
  • Recycling
  • UBAP1
  • Vesicular pH

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