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Histone modification dynamics as revealed by a multicolor immunofluorescence-based single-cell analysis

View ORCID ProfileYoko Hayashi-Takanaka, Yuto Kina, Fumiaki Nakamura, Leontine E. Becking, Yoichi Nakao, Takahiro Nagase, Naohito Nozaki, View ORCID ProfileHiroshi Kimura
doi: https://doi.org/10.1101/2020.01.01.892299
Yoko Hayashi-Takanaka
1Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan
2Graduate School of Frontier Biosciences, Osaka University, 1-3, Yamadaoka, Suita, 565-0871, Japan
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  • ORCID record for Yoko Hayashi-Takanaka
  • For correspondence: ythayashi@fbs.osaka-u.ac.jp hkimura@bio.titech.ac.jp
Yuto Kina
3Graduate School of Advanced Science and Engineering, Waseda University, Tokyo 169-8555, Japan
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Fumiaki Nakamura
3Graduate School of Advanced Science and Engineering, Waseda University, Tokyo 169-8555, Japan
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Leontine E. Becking
4Marine Animal Ecology Group, Wageningen University & Research, PO Box 338, Bode 36, 6700 AH Wageningen, The Netherlands
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Yoichi Nakao
3Graduate School of Advanced Science and Engineering, Waseda University, Tokyo 169-8555, Japan
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Takahiro Nagase
5Kazusa DNA Research Institute, Chiba, 292-0818, Japan
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Naohito Nozaki
6Mab Institute Inc, 2070-11, Osegi, Iida, 395-0157, Japan
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Hiroshi Kimura
1Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan
7Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan
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  • ORCID record for Hiroshi Kimura
  • For correspondence: ythayashi@fbs.osaka-u.ac.jp hkimura@bio.titech.ac.jp
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Abstract

Post-translational modifications on histones can be stable epigenetic marks and transient signals that can occur in response to internal and external stimuli. Levels of histone modifications fluctuate during the cell cycle and vary among different cell types. Here we describe a simple system to monitor the levels of multiple histone modifications in single cells by multicolor immunofluorescence using directly labeled modification-specific antibodies. We first analyzed histone H3 and H4 modifications during the cell cycle. Levels of active marks, such as acetylation and H3K4 methylation, were increased during the S phase, in association with chromatin duplication. By contrast, levels of some repressive modifications gradually increased during the G2 and the next G1 phases. We applied this method to validate the target modifications of various histone demethylases in cells using a transient overexpression system. We also screened chemical compounds in marine organism extracts that affect histone modifications and identified psammaplin A, which was previously reported to inhibit histone deacetylases. Thus, the method presented here is a powerful and convenient tool for analyzing the changes in histone modifications.

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Posted January 01, 2020.
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Histone modification dynamics as revealed by a multicolor immunofluorescence-based single-cell analysis
Yoko Hayashi-Takanaka, Yuto Kina, Fumiaki Nakamura, Leontine E. Becking, Yoichi Nakao, Takahiro Nagase, Naohito Nozaki, Hiroshi Kimura
bioRxiv 2020.01.01.892299; doi: https://doi.org/10.1101/2020.01.01.892299
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Histone modification dynamics as revealed by a multicolor immunofluorescence-based single-cell analysis
Yoko Hayashi-Takanaka, Yuto Kina, Fumiaki Nakamura, Leontine E. Becking, Yoichi Nakao, Takahiro Nagase, Naohito Nozaki, Hiroshi Kimura
bioRxiv 2020.01.01.892299; doi: https://doi.org/10.1101/2020.01.01.892299

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