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Mol Biol Cell. 2017 Aug 15;28(17):2290-2302. doi: 10.1091/mbc.E16-12-0870. Epub 2017 Jun 14.

HiHiMap: single-cell quantitation of histones and histone posttranslational modifications across the cell cycle by high-throughput imaging.

Author information

1
Cell Biology of Genomes, National Institutes of Health, Bethesda, MD 20892.
2
NCI High-Throughput Imaging Facility, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
3
Cell Biology of Genomes, National Institutes of Health, Bethesda, MD 20892 mistelit@mail.nih.gov.

Abstract

We describe High-throughput Histone Mapping (HiHiMap), a high-throughput imaging method to measure histones and histone posttranslational modifications (PTMs) in single cells. HiHiMap uses imaging-based quantification of DNA and cyclin A to stage individual cells in the cell cycle to determine the levels of histones or histone PTMs in each stage of the cell cycle. As proof of principle, we apply HiHiMap to measure the level of 21 core histones, histone variants, and PTMs in primary, immortalized, and transformed cells. We identify several histone modifications associated with oncogenic transformation. HiHiMap allows the rapid, high-throughput study of histones and histone PTMs across the cell cycle and the study of subpopulations of cells.

PMID:
28615324
PMCID:
PMC5555657
DOI:
10.1091/mbc.E16-12-0870
[Indexed for MEDLINE]
Free PMC Article

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