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Analysis of Nucleocytoplasmic Protein Shuttling by Imaging Flow Cytometry

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Imaging Flow Cytometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1389))

Abstract

Many intracellular signal transduction events involve the reversible shuttling of proteins between the cytoplasm and the nucleus. Study of these processes requires imaging information on the protein localization in a given cell and a large number of measurements to obtain sufficient statistics on the protein localization in the whole population. The protocol describes method for quantitative imaging flow cytometry analysis of intracellular distribution of NF-kappaB in ARPE-19 cells stained with specific fluorochrome-conjugated antibodies. The described technique alone or in combination with standard flow cytometry methods can be applied to study any protein undergoing translocation from cytoplasm into the nucleus in a variety of cell lines as well as in heterogeneous primary cell populations.

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Acknowledgements

The work was supported in part by NIH S10 RR023459 grant, Harvard Pilot grant, the Russian Foundation for Basic Research grants and 13-04-40189-H, Swiss IBD grant and PI Nuris. We are also grateful to Aleksandra Gorelova (Harvard University) for help with the editing and preparation of the manuscript.

Author information

Authors and Affiliations

  1. Department of Biomedicine, University of Basel and University Hospital Basel, CH-4031, Basel, Switzerland

    Elizaveta Fasler-Kan

  2. Department of Pediatric Surgery and Department of Clinical Research, Inselspital University Hospital, University of Bern, Bern, CH-3010, Switzerland

    Elizaveta Fasler-Kan

  3. Nazarbayev University, Astana, Kazakhstan

    Yeldar Baiken & Natasha S. Barteneva

  4. Cellular and Molecular Medicine Program, Boston Children’s Hospital, Boston, MA, USA

    Yeldar Baiken

  5. Department of Cell Biology and Histology, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia

    Ivan A. Vorobjev

  6. A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia

    Ivan A. Vorobjev

  7. Department of Pediatrics, Harvard Medical School, Boston, MA, USA

    Natasha S. Barteneva

Authors
  1. Elizaveta Fasler-Kan
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  2. Yeldar Baiken
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  3. Ivan A. Vorobjev
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  4. Natasha S. Barteneva
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Corresponding author

Correspondence to Elizaveta Fasler-Kan .

Editor information

Editors and Affiliations

  1. Cellular and Molecular Medicine Program, Boston Children’s Hospital, Boston, Massachusetts, USA

    Natasha S. Barteneva

  2. A.N. Belozersky Institute of Physico-Che, M.V. Lomonosov Moscow State University, Moscow, Russia

    Ivan A. Vorobjev

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Fasler-Kan, E., Baiken, Y., Vorobjev, I.A., Barteneva, N.S. (2016). Analysis of Nucleocytoplasmic Protein Shuttling by Imaging Flow Cytometry. In: Barteneva, N., Vorobjev, I. (eds) Imaging Flow Cytometry. Methods in Molecular Biology, vol 1389. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3302-0_8

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  • DOI: https://doi.org/10.1007/978-1-4939-3302-0_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3300-6

  • Online ISBN: 978-1-4939-3302-0

  • eBook Packages: Springer Protocols

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