FIG. 10.

HIV-1 binding to PM1 T cells triggers raft aggregation and cocapping of HIV-1 receptors. PM1 cells were incubated on ice for 30 min without (top row) or with (middle three rows) purified HIV-1 NL4-3 (5 cpm of RT/cell) and then incubated at 37°C for 45 min. (Bottom row) Alternatively, PM1 cells were first treated with 10 mM MβCD to deplete cholesterol and then infected with NL4-3. The cells were fixed with 4% paraformaldehyde and analyzed for the surface distribution of HIV-1 gp120, CCR5, the raft marker GM1, or the nonraft marker CD71. HIV-1 gp120 was visualized using FITC-conjugated monoclonal anti-gp120 (middle row) or mouse monoclonal anti-gp120 and goat anti-mouse antibodies labeled with Alexa Fluor 594 (bottom two rows). CCR5 was visualized with FITC-conjugated monoclonal anti-CCR5 (2D7) antibody. GM1 was labeled with Alexa Fluor 594 conjugates of CT-B, and CD71 was labeled with monoclonal FITC-conjugated anti-CD71 antibody.