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Conic, S.* ; Desplancq, D.* ; Ferrand, A.* ; Fischer, V.* ; Heyer, V.* ; Martin, B.R.S.* ; Pontabry, J. ; Oulad-Abdelghani, M.* ; Babu, K.N.* ; Wright, G.D.* ; Molina, N.* ; Weiss, E.* ; Tora, L.*

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

J. Cell Biol. 217, 1537-1552 (2018)

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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 (VAN IMA) 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, VAN IMA 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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Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Rna-polymerase-ii; Structured Illumination Microscopy; Tata-binding Protein; Living Cells; Monoclonal-antibodies; Mammalian-cells; Superresolution Microscopy; Cancer-cells; Dna-damage; Dynamics

ISSN (print) / ISBN 0021-9525

e-ISSN 1540-8140

Journal Journal of Cell Biology, The

Quellenangaben Volume: 217, Issue: 4, Pages: 1537-1552

Publisher Rockefeller University Press

Publishing Place New York

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Epigenetics and Stem Cells (IES)