Wang, Lili; Sandmeyer, Alice; Hübner, Wolfgang ; Li, Hongru; Huser, Thomas ; Chen, Benjamin K.: A Replication-Competent HIV Clone Carrying GFP-Env Reveals Rapid Env Recycling at the HIV-1 T Cell Virological Synapse. In: Viruses. Jg.14 H. 1. 2022
Inhalt
- Materials and Methods
- Cell Lines and Virus Production
- Human Primary CD4 T Cells
- Viruses
- p24 ELISA
- Western Blot Analysis
- Infectivity TZM-bl Assay
- Cell-to-Cell Transfer Assay
- Growth Curve of HIV Env-V4.2-sfGFP
- Fluorescence Microscopy Sample Preparation
- Confocal and Live Imaging
- Fluorescence Recovery after Photobleaching (FRAP)
- Super-Resolution Optical Microscopy of HIV-Infected T Cells
- Results
- Engineering an Infectious HIV Carrying a sfGFP Insertion into the Env V4 or V5 Domains
- Imaging HIV-1 Carrying Fluorescent Env Constructs
- A dual Fluorescent Protein-Expressing HIV with Gag-iCherry and Env-sfGFP Participates in VS-Mediated HIV Transfer
- Method for Pulse-Chase Labeling of Surface Env Tracks Endocytosis and Relocalization to the VS
- Fluorescence Recovery after Photobleaching (FRAP) of HIV Env V4.2 sfGFP-Gag-iCherry at VS Reveals Constitutive Turnover of Env near the VS
- High Turnover of Env Proximal to the VS Requires Endocytosis of Env Using a Membrane Proximal Tyrosine Y712
- Discussion
- References