Abstract
Nucleocytoplasmic transport is crucial not only for basic cellular activities but also for the physiological adaptation of cells to various environmental stimuli that affect development, cell-fate determination, or disease development. The basic transport mechanisms have been revealed during the past two decades through the identification and biochemical characterizations of factors mediating the transport, dissecting the transport process and examining the function of nuclear pore complexes (NPCs). In this chapter, we describe methods for a nuclear transport reconstitution assay using digitonin-permeabilized mammalian cells. The transport assay can be generally conducted in the lab without special equipment. The assay system is efficient and significantly contributes to the study of nucleocytoplasmic transport.
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Acknowledgments
We are grateful to the members of the Cellular Dynamics Laboratory. Our work was supported by RIKEN Special Project Funding for Basic Science in Cellular System Project Research, the Japan Society for the Promotion of Science (JSPS) through the “Funding Program for Next Generation World-Leading Researchers (NEXT Program),” initiated by the Council for Science and Technology Policy (CSTP) to N.I., and MEXT grants-in-aid to S.K. and T.F.
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Kose, S., Funakoshi, T., Imamoto, N. (2015). Reconstitution of Nucleocytoplasmic Transport Using Digitonin-Permeabilized Cells. In: Nakagawa, S., Hirose, T. (eds) Nuclear Bodies and Noncoding RNAs. Methods in Molecular Biology, vol 1262. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2253-6_18
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DOI: https://doi.org/10.1007/978-1-4939-2253-6_18
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