Abstract
This chapter describes a method used to assay the cell cycle dynamics of the centromeric histone H3 variant CENP-A in meiosis using Drosophila males as the experimental system. Specifically, we describe a method that combines Immunofluorescence (IF) and Fluorescence in-situ Hybridization (FISH) protocols, performed on fixed Drosophila testes. An advantage of this protocol is the ability to localize individual centromeres on the four Drosophila homologous chromosomes that form distinct nuclear territories in spermatocytes. We also describe a method to quantify CENP-A focal intensities using Image J software.
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References
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Acknowledgments
This work is supported by a Science Foundation Ireland (SFI)-Health Research Board-Wellcome Trust Research Career Development Fellowship and SFI-President of Ireland Young Researcher Award (PIYRA) to EMD. CMC is funded by a National University of Ireland Galway, College of Science Scholarship and SFI-PIYRA.
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Collins, C.M., Dunleavy, E.M. (2018). Imaging and Quantitation of Assembly Dynamics of the Centromeric Histone H3 Variant CENP-A in Drosophila melanogaster Spermatocytes by Immunofluorescence and Fluorescence In-Situ Hybridization (Immuno-FISH). In: Orsi, G., Almouzni, G. (eds) Histone Variants. Methods in Molecular Biology, vol 1832. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8663-7_18
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DOI: https://doi.org/10.1007/978-1-4939-8663-7_18
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