Abstract
Mammalian pre-implantation embryos represent a highly dynamic experimental model for comparative studies of nuclear structure and functions in the context of gradual reactivation of transcription. Here, we present details of the methods that allow localizing RNA polymerase II in mouse pre-implantation embryos with specific antibodies, using fluorescent/confocal and electron microscopy. We stress the special aspects of immunolabeling protocols in respect to the embryonic material. We made a special emphasis on the essential steps preceding the immunocytochemical experiments. In particular, we consider the procedures of female hormonal stimulation and embryo collection. The described approaches are also applicable to study other nuclear proteins.
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Acknowledgment
This work was supported by the granting program “Molecular and Cell Biology” of the Russian Academy of Sciences and the Russian Foundation for Basic Research (grant No. 15-04-01857).
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Bogolyubova, I.O., Bogolyubov, D.S. (2017). Detection of RNA Polymerase II in Mouse Embryos During Zygotic Genome Activation Using Immunocytochemistry. In: Lee, K. (eds) Zygotic Genome Activation. Methods in Molecular Biology, vol 1605. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6988-3_10
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DOI: https://doi.org/10.1007/978-1-4939-6988-3_10
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