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Quantitative Analysis of PcG-Associated Condensates by Stochastic Optical Reconstruction Microscopy (STORM)

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Polycomb Group Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2655))

Abstract

The polycomb group (PcG) proteins play a central role in the maintenance of a repressive state of gene expression. Recent findings demonstrate that PcG components are organized into nuclear condensates, contributing to the resha** of chromatin architecture in physiological and pathological conditions, thus affecting the nuclear mechanics. In this context, direct stochastic optical reconstruction microscopy (dSTORM) provides an effective tool to achieve a detailed characterization of PcG condensates by visualizing them at a nanometric level. Furthermore, by analyzing dSTORM datasets with cluster analysis algorithms, quantitative information can be yielded regarding protein numbers, grou**, and spatial organization. Here, we describe how to set up a dSTORM experiment and perform the data analysis to study PcG complexes’ components in adhesion cells quantitatively.

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Author information

Authors and Affiliations

  1. Chromatin Biology & Epigenetics Lab, Department of Cellular, Computational, and Integrative Biology (CIBIO), University of Trento, Trento, Italy

    Silvia Scalisi, Sarah D’Annunzio & Alessio Zippo

  2. Laboratoire Angevin de Recherche en Ingénierie des Systèmes, UMR INRAE IRHS, Université d’Angers, Angers, France

    Ali Ahmad & David Rousseau

Authors
  1. Silvia Scalisi
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  2. Ali Ahmad
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  3. Sarah D’Annunzio
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  4. David Rousseau
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  5. Alessio Zippo
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Corresponding author

Correspondence to Alessio Zippo .

Editor information

Editors and Affiliations

  1. Institute of Biomedical Technologies, National Research Council, Milan, Italy

    Chiara Lanzuolo

  2. Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, INGM, Milan, Italy

    Federica Marasca

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Scalisi, S., Ahmad, A., D’Annunzio, S., Rousseau, D., Zippo, A. (2023). Quantitative Analysis of PcG-Associated Condensates by Stochastic Optical Reconstruction Microscopy (STORM). In: Lanzuolo, C., Marasca, F. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 2655. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3143-0_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3143-0_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3142-3

  • Online ISBN: 978-1-0716-3143-0

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