Abstract
Epigenetic mechanisms lead to the stable regulation of gene expression without alteration of DNA and trigger initiation and/or maintenance of cell-type-specific transcriptional profiles. Indeed, modulation of chromatin structure and the global 3D organization of the genome and nuclear architecture participate in the precise control of transcription. Thus, dissection of these epigenetic mechanisms is essential for our understanding of gene regulation. In this chapter, we describe challenging combinations of immunofluorescence, and RNA and DNA fluorescent in situ hybridization and their application to our studies of a remarkable example of epigenetic control of gene expression in female mammals, the process of X chromosome inactivation.
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Acknowledgments
We thank the EU Network of Excellence “The Epigenome” for permission to republish these procedures. Funding sources for this work were the Human Frontiers Science Program, EU FP6 Integrated Project HEROIC (LSHG-CT-2005-018883), the Schlumberger Foundation for Research, and the EU FP6 Network of Excellence “The Epigenome” (LSHG-CT-2004-503433). JC was funded by the French “Ligue Nationale contre le Cancer” and the HEROIC IP; SA was funded by the French “Centre National de la Recherche Scientifique”; JCC was funded by the HEROIC IP.
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Chaumeil, J., Augui, S., Chow, J.C., Heard, E. (2008). Combined Immunofluorescence, RNA Fluorescent In Situ Hybridization, and DNA Fluorescent In Situ Hybridization to Study Chromatin Changes, Transcriptional Activity, Nuclear Organization, and X-Chromosome Inactivation. In: Hancock, R. (eds) The Nucleus. Methods in Molecular Biology, vol 463. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-406-3_18
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DOI: https://doi.org/10.1007/978-1-59745-406-3_18
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-977-2
Online ISBN: 978-1-59745-406-3
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