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1Department of Pathology,New York University Langone School of Medicine, 2Program in Molecular Pathogenesis,Marty and Helen Kimmel Center for Biology and Medicine and Skirball Institute for Biomolecular Medicine, 3Laboratory of Molecular Immunogenetics, National Institute of Arthritis and Musculoskeletal and Skin Diseases,National Institutes of Health, 4Veteran Affairs New York Harbor Healthcare System
Chapters
- 00:05Title
- 02:18Labeling GP120
- 03:52Flow Cell Assembly and Bilayer Preparation: His6-tagged GP120 DH12
- 06:11Injection of Cells and Immunofluorescence Staining
- 07:49Data Analysis
- 08:56Results: Imaging Lck and Fyn at HIV-1 Envelope-induced Virological Synapse
- 10:53Conclusion
This article describes a method to visualize formation of an HIV-1 envelope-induced virological synapse on glass supported planar bilayers by total internal reflection fluorescence (TIRF) microscopy. The method can also be combined with immunofluorescence staining to detect activation and redistribution of signaling molecules that occur during HIV-1 envelope-induced virological synapse formation.
Tags
HIV-1EnvelopeVirological SynapseSignalingSynthetic Lipid BilayersCell To Cell TransmissionCD4+ T CellsF-actinGp120Chemokine Receptor (CKR)CCR5CXCR4LFA-1ICAM FamilyVirus-infected Donor CellsTarget CellsHIV-1 VirionsIntracellular Signaling EventsTCR SignalosomeLckCD3ζZAP70LATSLP-76ItkPLCγSupramolecular Arrangement
