Imaging of native transcription factors and histone phosphorylation at high resolution in live cells
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...
| Main Authors: | , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2018
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| Online Access: | https://hdl.handle.net/10356/88771 http://hdl.handle.net/10220/44728 |
| _version_ | 1811688762999046144 |
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| author | Conic, Sascha Desplancq, Dominique Ferrand, Alexia Fischer, Veronique Heyer, Vincent Reina San Martin, Bernardo Pontabry, Julien Oulad-Abdelghani, Mustapha Babu N., Kishore Wright, Graham D. Molina, Nacho Weiss, Etienne Tora, László |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Conic, Sascha Desplancq, Dominique Ferrand, Alexia Fischer, Veronique Heyer, Vincent Reina San Martin, Bernardo Pontabry, Julien Oulad-Abdelghani, Mustapha Babu N., Kishore Wright, Graham D. Molina, Nacho Weiss, Etienne Tora, László |
| author_sort | Conic, Sascha |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T05:37:22Z |
| format | Journal Article |
| id | ntu-10356/88771 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:37:22Z |
| publishDate | 2018 |
| record_format | dspace |
| spelling | ntu-10356/887712023-02-28T17:02:47Z Imaging of native transcription factors and histone phosphorylation at high resolution in live cells Conic, Sascha Desplancq, Dominique Ferrand, Alexia Fischer, Veronique Heyer, Vincent Reina San Martin, Bernardo Pontabry, Julien Oulad-Abdelghani, Mustapha Babu N., Kishore Wright, Graham D. Molina, Nacho Weiss, Etienne Tora, László School of Biological Sciences Transcription Histone 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. ASTAR (Agency for Sci., Tech. and Research, S’pore) Published version 2018-04-26T06:40:20Z 2019-12-06T17:10:36Z 2018-04-26T06:40:20Z 2019-12-06T17:10:36Z 2018 Journal Article Conic, S., Desplancq, D., Ferrand, A., Fischer, V., Heyer, V., Reina San Martin, B., et al. (2018). Imaging of native transcription factors and histone phosphorylation at high resolution in live cells. Journal of Cell Biology, 217(4), 1537-1552. 0021-9525 https://hdl.handle.net/10356/88771 http://hdl.handle.net/10220/44728 10.1083/jcb.201709153 en Journal of Cell Biology © 2018 Conic et al. 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/). 16 p. application/pdf |
| spellingShingle | Transcription Histone Conic, Sascha Desplancq, Dominique Ferrand, Alexia Fischer, Veronique Heyer, Vincent Reina San Martin, Bernardo Pontabry, Julien Oulad-Abdelghani, Mustapha Babu N., Kishore Wright, Graham D. Molina, Nacho Weiss, Etienne Tora, László Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title | Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title_full | Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title_fullStr | Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title_full_unstemmed | Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title_short | Imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| title_sort | imaging of native transcription factors and histone phosphorylation at high resolution in live cells |
| topic | Transcription Histone |
| url | https://hdl.handle.net/10356/88771 http://hdl.handle.net/10220/44728 |
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