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Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction–Associated Polarity Proteins

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Cell Polarity Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2438))

Abstract

Here, we present a detailed protocol for fluorescence recovery after photobleaching (FRAP) to measure the dynamics of junctional populations of proteins in living tissue. Specifically, we describe how to perform FRAP in Drosophila pupal wings on fluorescently tagged core planar polarity proteins, which exhibit relatively slow junctional turnover. We provide a step-by-step practical guide to performing FRAP, and list a series of controls and optimizations to do before conducting a FRAP experiment. Finally, we describe how to present the FRAP data for publication.

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Authors and Affiliations

  1. School of Biosciences, University of Sheffield, Sheffield, UK

    Samantha J. Warrington, Helen Strutt & David Strutt

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  1. Samantha J. Warrington
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  2. Helen Strutt
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  3. David Strutt
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Correspondence to David Strutt .

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Editors and Affiliations

  1. Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    Chenbei Chang

  2. Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    Jianbo Wang

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Warrington, S.J., Strutt, H., Strutt, D. (2022). Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction–Associated Polarity Proteins. In: Chang, C., Wang, J. (eds) Cell Polarity Signaling. Methods in Molecular Biology, vol 2438. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2035-9_1

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  • DOI: https://doi.org/10.1007/978-1-0716-2035-9_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2034-2

  • Online ISBN: 978-1-0716-2035-9

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