Abstract
The spatial distribution of genes in the nucleus emerges as an important factor in gene regulation and epigenetics. The position of loci relative to each other, to nuclear landmarks such as the nucleolus and chromocenters, as well as to chromatin proteins is therefore highly interesting. With fluorescent in situ hybridization (FISH) specific DNA sequences can be stained and antibodies allow the detection of specific proteins. Here, we present two protocols that preserve the 3D structure of nuclei. With whole-mount FISH, specific sequences can be stained in intact tissues and, secondly, a combined immunolabeling and FISH protocol on acrylamide-embedded nuclei makes it possible to stain DNA sequences and proteins simultaneously.
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Acknowledgments
T.B. was financially supported by the European Commission Seventh Framework-People-2012-ITN (Project EpiTRAITS, GA-316965) and M.K. was financially supported by the Netherlands’ Organization for Scientific Research (ALW2PJ/09053).
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Bey, T.D., Koini, M., Fransz, P. (2018). Fluorescence In Situ Hybridization (FISH) and Immunolabeling on 3D Preserved Nuclei. In: Bemer, M., Baroux, C. (eds) Plant Chromatin Dynamics. Methods in Molecular Biology, vol 1675. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7318-7_27
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DOI: https://doi.org/10.1007/978-1-4939-7318-7_27
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7317-0
Online ISBN: 978-1-4939-7318-7
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