| Title |
Fig. 3. Trimer formation by HIV-1 Env protein expressed by VVEnv-22. Hep2 cells were infected with the virus at different multiplicities of infection (MOI): 0.1 (lane 1), 0.2 (lane 2), 0.5 (lanes 3, 5) PFU/cell. Negative control – uninfected Hep2 cells (lane 4). After 48 h, the cells were removed and the plasma membrane fraction containing HIV-1 proteins was purified. Samples were dissolved in sample loading buffer without reducing agents at room temperature (lanes 1‒3) or with reducing agents (200 mM dithiothreitol, 200 mM β-mercaptoethanol, 8 M urea) for 5 min at 96°C (lane 5). Proteins were separated in 4–15% SDS-PAAG and analyzed by immunoblotting. |