Abstract
Super-resolution confocal live imaging microscopy (SCLIM) we developed provides high-speed, high-resolution, three- and four-dimensional, and multicolor simultaneous imaging. Using this technology, we are now able to observe the fine details of various dynamic events going on in living cells, such as membrane traffic and organelle dynamics. The retention using selective hooks (RUSH) system is a powerful tool to control synchronous release of natural cargo proteins of interest from the endoplasmic reticulum in mammalian cells. In this chapter, we describe a method for visualizing secretory cargo traffic within and around the Golgi apparatus in HeLa cells using SCLIM in combination with the RUSH assay.
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Change history
11 February 2023
The original version of the book was inadvertently published without incorporating the author’s proof corrections mentioned below. The chapters have now been corrected and approved by the author.
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Acknowledgments
We are grateful to Kumiko Ishii for technical assistance in sample preparation and microscopic observation. We also thank all the members of the Nakano laboratory for helpful discussions. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan (grant numbers 17H05756, 19K06669, and 19H04764 to T.T.; 17H06420 and 18H05275 to A.N.) and Japan Science and Technology Agency (JST) CREST program (grant number JPMJCR21E3 to T.T.). D.M. is supported by the RIKEN Special Postdoctoral Researcher Program.
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This article is dedicated to Yasuhito Kosugi, who passed away in 2020.
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Tojima, T., Miyashiro, D., Kosugi, Y., Nakano, A. (2023). Super-Resolution Live Imaging of Cargo Traffic Through the Golgi Apparatus in Mammalian Cells. In: Wang, Y., Lupashin, V.V., Graham, T.R. (eds) Golgi. Methods in Molecular Biology, vol 2557. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2639-9_10
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DOI: https://doi.org/10.1007/978-1-0716-2639-9_10
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