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

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Book cover The ESCRT Complexes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1998))

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.

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Correspondence to Pirjo M. Apaja or Arnim Pause .

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Kazan, J.M., Lukacs, G.L., Apaja, P.M., Pause, A. (2019). Single Cell Fluorescence Ratio Image Analysis for Studying ESCRT Function in Receptor Trafficking. In: Culetto, E., Legouis, R. (eds) The ESCRT Complexes. Methods in Molecular Biology, vol 1998. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9492-2_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9492-2_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9491-5

  • Online ISBN: 978-1-4939-9492-2

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