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J Cell Biol. 2018 Apr 2;217(4):1537-1552. doi: 10.1083/jcb.201709153. Epub 2018 Feb 12.

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

Conic S1,2,3,4, Desplancq D5, Ferrand A6, Fischer V1,2,3,4, Heyer V1,2,3,4, Reina San Martin B1,2,3,4, Pontabry J1,2,3,4,7, Oulad-Abdelghani M1,2,3,4, Babu N K8, Wright GD9, Molina N1,2,3,4, Weiss E10, Tora L11,2,3,4,8.

Author information

1
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France.
2
Centre National de la Recherche Scientifique, UMR7104, Illkirch, France.
3
Institut National de la Santé et de la Recherche Médicale, U964, Illkirch, France.
4
Université de Strasbourg, Illkirch, France.
5
Institut de Recherche de l'ESBS, UMR 7242, Illkirch, France.
6
Imaging Core Facility, Biozentrum, University of Basel, Basel, Switzerland.
7
Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Institute of Epigenetics and Stem Cells, München, Germany.
8
School of Biological Sciences, Nanyang Technological University, Singapore.
9
Institute of Medical Biology, A*STAR, Singapore, Singapore.
10
Institut de Recherche de l'ESBS, UMR 7242, Illkirch, France etienne.weiss@unistra.fr.
11
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France laszlo@igbmc.fr.

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.

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