Abstract
Vesicle endocytosis at the plasma membrane is associated with a precise temporal choreography in the recruitment of cytosolic proteins that sense, generate, or stabilize locally curved membrane regions. To dissect the role of membrane curvature sensing from other co-occurring events during the initial steps of endocytosis, we developed a method to artificially induce nanoscale deformations of the PM in living cells that is based on cone-shaped nanostructures (i.e., Nanocones). When cultured on Nanocones, cells create stable inward plasma membrane deformations to which curvature-sensing proteins are recruited. Here, we provide a detailed protocol how to use Nanocones to study recruitment during the initial steps of endocytosis in cells by fluorescence and electron microscopy.
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Acknowledgments
S. J. acknowledges support from the Korea Foundation for Advanced Studies (KFAS) for graduate fellowship. M. G. was supported by fellowships from the Swiss National Science Foundation (No. PBBSP3-123159), Novartis Jubilaeumsstiftung, Stanford Deans Postdoctoral Fellowship and grants from the German Research Foundation (Cluster of Excellence EXC 1003, Cells in Motion, CiM, Münster, Germany).
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Jeong, S., Galic, M. (2014). Nanocones to Study Initial Steps of Endocytosis. In: Ivanov, A. (eds) Exocytosis and Endocytosis. Methods in Molecular Biology, vol 1174. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0944-5_19
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DOI: https://doi.org/10.1007/978-1-4939-0944-5_19
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0943-8
Online ISBN: 978-1-4939-0944-5
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