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3D Time-Lapse Analysis of Rab11/FIP5 Complex: Spatiotemporal Dynamics During Apical Lumen Formation

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Rab GTPases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1298))

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Abstract

Fluorescent imaging of fixed cells grown in two-dimensional (2D) cultures is one of the most widely used techniques for observing protein localization and distribution within cells. Although this technique can also be applied to polarized epithelial cells that form three-dimensional (3D) cysts when grown in a Matrigel matrix suspension, there are still significant limitations in imaging cells fixed at a particular point in time. Here, we describe the use of 3D time-lapse imaging of live cells to observe the dynamics of apical membrane initiation site (AMIS) formation and lumen expansion in polarized epithelial cells.

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Acknowledgement

This work was supported by NIH grant R01-DK064380 to R.P. and NIH Pre-doctoral Training Program T32-GM08730.

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Correspondence to Rytis Prekeris .

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Mangan, A., Prekeris, R. (2015). 3D Time-Lapse Analysis of Rab11/FIP5 Complex: Spatiotemporal Dynamics During Apical Lumen Formation. In: Li, G. (eds) Rab GTPases. Methods in Molecular Biology, vol 1298. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2569-8_15

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  • DOI: https://doi.org/10.1007/978-1-4939-2569-8_15

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2568-1

  • Online ISBN: 978-1-4939-2569-8

  • eBook Packages: Springer Protocols

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