Abstract
Fluorescence microscopy and fluorescent protein (FP)-tagged GLUT4 molecule have been great tools to characterize GLUT4 localization and dynamics inside the cell. However, it was difficult to distinguish GLUT4 storage vesicles (GSVs) from other intracellular compartments containing GLUT4 in live cells. Here, we describe the use of IRAP-pHluorin and total internal reflection fluorescence (TIRF) microscopy to selectively visualize GSVs and Rab proteins that associate with GSVs. This assay is also valuable to further defining GSV identity by unraveling other GSV-associated proteins.
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Acknowledgement
We thank all members of J.L.-S.’s group for helpful discussions.
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Chen, Y., Lippincott-Schwartz, J. (2015). Selective Visualization of GLUT4 Storage Vesicles and Associated Rab Proteins Using IRAP-pHluorin. 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_14
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DOI: https://doi.org/10.1007/978-1-4939-2569-8_14
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2568-1
Online ISBN: 978-1-4939-2569-8
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