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Imaging of native transcription factors and histone phosphorylation at high resolution in live cells

Sascha Conic, Dominique Desplancq, View ORCID ProfileAlexia Ferrand, Veronique Fischer, Vincent Heyer, Bernardo Reina San Martin, Julien Pontabry, Mustapha Oulad-Abdelghani, Kishore Babu N., View ORCID ProfileGraham D. Wright, Nacho Molina, Etienne Weiss  Correspondence email, View ORCID ProfileLászló Tora  Correspondence email
Sascha Conic
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Dominique Desplancq
Institut de Recherche de l’ESBS, UMR 7242, Illkirch, France
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Alexia Ferrand
Imaging Core Facility, Biozentrum, University of Basel, Basel, Switzerland
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  • ORCID record for Alexia Ferrand
Veronique Fischer
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Vincent Heyer
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Bernardo Reina San Martin
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Julien Pontabry
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, FranceHelmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Institute of Epigenetics and Stem Cells, München, Germany
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Mustapha Oulad-Abdelghani
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Kishore Babu N.
School of Biological Sciences, Nanyang Technological University, Singapore
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Graham D. Wright
Institute of Medical Biology, A*STAR, Singapore, Singapore
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  • ORCID record for Graham D. Wright
Nacho Molina
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Etienne Weiss
Institut de Recherche de l’ESBS, UMR 7242, Illkirch, France
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  • For correspondence: etienne.weiss@unistra.fr
László Tora
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, FranceSchool of Biological Sciences, Nanyang Technological University, Singapore
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  • ORCID record for László Tora
  • For correspondence: laszlo@igbmc.fr
DOI: 10.1083/jcb.201709153 | Published February 12, 2018
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Abstract

Fluorescent labeling of endogenous proteins for live-cell imaging without exogenous expression of tagged proteins or genetic manipulations has not been routinely possible. We describe a simple versatile antibody-based imaging approach (VANIMA) for the precise localization and tracking of endogenous nuclear factors. Our protocol can be implemented in every laboratory allowing the efficient and nonharmful delivery of organic dye-conjugated antibodies, or antibody fragments, into different metazoan cell types. Live-cell imaging permits following the labeled probes bound to their endogenous targets. By using conventional and super-resolution imaging we show dynamic changes in the distribution of several nuclear transcription factors (i.e., RNA polymerase II or TAF10), and specific phosphorylated histones (γH2AX), upon distinct biological stimuli at the nanometer scale. Hence, considering the large panel of available antibodies and the simplicity of their implementation, VANIMA can be used to uncover novel biological information based on the dynamic behavior of transcription factors or posttranslational modifications in the nucleus of single live cells.

  • Submitted: 28 September 2017
  • Revision received 15 December 2017
  • Accepted: 18 January 2018
Creative Commons logoCreative Commons logohttp://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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© 2018 Conic et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Imaging of native transcription factors and histone phosphorylation at high resolution in live cells
Sascha Conic, Dominique Desplancq, Alexia Ferrand, Veronique Fischer, Vincent Heyer, Bernardo Reina San Martin, Julien Pontabry, Mustapha Oulad-Abdelghani, Kishore Babu N., Graham D. Wright, Nacho Molina, Etienne Weiss, László Tora
J Cell Biol Feb 2018, jcb.201709153; DOI: 10.1083/jcb.201709153

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The Journal of Cell Biology: 217 (4)

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April 2, 2018
Volume 217, No. 4

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