Abstract
Proximity ligation assay (PLA) is a newly developed technique that outperforms the traditional immunoassays for visualizing the in situ endogenous protein-protein interactions and localizations and the activation of proteins in cell culture systems as well as in tissue sections. PLA, when combined with cellular marker staining, becomes a powerful approach to identify differential interaction of the proteins of endosomal sorting complex required for transport (ESCRT) at distinct stages of virus infection. In this chapter, we describe a PLA protocol to study the localization and interaction between the ESCRT protein TSG101 and endosomal markers in early stages of viral endocytosis in in vitro infected cells.
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Acknowledgments
This study was supported in part by the Public Health Service grants CA 168472 and CA 180758, RFUMS H. M. Bligh Cancer Research Fund, and USF start up fund to Bala Chandran.
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Kumar, B., Veettil, M.V., Roy, A., Chandran, B. (2019). Proximity Ligation Assay (PLA) to Determine the Endosomal Localization of ESCRT Subunit in Virus-Infected Cells. In: Culetto, E., Legouis, R. (eds) The ESCRT Complexes. Methods in Molecular Biology, vol 1998. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9492-2_5
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DOI: https://doi.org/10.1007/978-1-4939-9492-2_5
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