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Using Bimolecular Fluorescence Complementation (BiFC) with Photoactivated Localization Microscopy (PALM) to Analyze Hox/Cofactor Interactions at the Super Resolution Scale in Drosophila Salivary Glands

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HOX Genes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2889))

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Abstract

Bimolecular Fluorescence Complementation (BiFC) is a powerful molecular imaging method used to visualize protein-protein interactions (PPIs) in living cells or organisms. BiFC is based on the reassociation of hemi-fragments of a monomeric fluorescent protein upon spatial proximity. It is compatible with conventional light microscopy, providing a resolution that is constrained by the diffraction of light to around 250 nm. PAmCherry1, a fluorescent protein compatible for BiFC and Photoactivated Localization Microscopy (PALM), allows the analysis of PPIs with nanometer spatial resolution and single molecule sensitivity. To date, this so-called BiFC-PALM approach has only been described in human cell culture. Here, we present a protocol for performing BiFC-PALM in Drosophila larval salivary glands, using the interaction between the Hox protein Ultrabithorax (Ubx) and the generic Hox cofactor Extradenticle (Exd) as a model system.

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Acknowledgements

This work was supported by Fondation pour la Recherche Médicale (FRM 160896), CNRS, ENS-Lyon and UAR3444/US8.

We thank the Bloomington stock center for the Drosophila fly lines and platforms of the UAR3444/US8 (Arthrotools and PLATIM). We thank Elodie Chatre for her help on the super resolution microscope and her critical reading of the manuscript.

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Authors and Affiliations

  1. Institut de Génomique Fonctionnelle de Lyon (IGFL), UMR5242, Ecole Normale Supérieure de Lyon (ENSL), CNRS, Université de Lyon, Lyon, France

    Solène Vanderperre, Marilyne Duffraisse & Samir Merabet

  2. FLPENS, CNRS UMR572, Ecole Normale Supérieure de Lyon (ENSL), Université de Lyon, Lyon, France

    Christophe Place

Authors
  1. Solène Vanderperre
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  2. Marilyne Duffraisse
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  3. Christophe Place
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  4. Samir Merabet
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Corresponding author

Correspondence to Samir Merabet .

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Editors and Affiliations

  1. Yale Center for Genome Analysis, Yale University, West Haven, CT, USA

    Bony De Kumar

  2. Yale Center for Genome Analysis, Yale University, West Haven, CT, USA

    Ashish Shelar

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Vanderperre, S., Duffraisse, M., Place, C., Merabet, S. (2025). Using Bimolecular Fluorescence Complementation (BiFC) with Photoactivated Localization Microscopy (PALM) to Analyze Hox/Cofactor Interactions at the Super Resolution Scale in Drosophila Salivary Glands. In: De Kumar, B., Shelar, A. (eds) HOX Genes. Methods in Molecular Biology, vol 2889. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-4322-8_4

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  • DOI: https://doi.org/10.1007/978-1-0716-4322-8_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-4321-1

  • Online ISBN: 978-1-0716-4322-8

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