Figure 1

Tetraspanins are not release factors. HeLa cells were transduced with the indicated shRNA lentiviral vectors at MOI = 8, then transfected with pNL4-3 provirus 48 hours post-transduction. 2 days later, cells and viruses were harvested as described in Methods, and analyzed by Western blot for CD63, CD9, CD81 and Gag levels (A), or p24 ELISA for p24 CA content (B). Normalized percent release was calculated by dividing the amount of p24 released by the total amount of p24 produced (cells + supernatant), then normalizing to the mock-treated sample (set to 100%). Data shown represent the mean of 4 independent experiments, +/- standard error of the mean (SEM). (C) HeLa cells were co-transfected with pNL4-3 and the indicated plasmids. Cells and viruses were harvested and analyzed by p24 ELISA, as in (B). Data shown represent the mean of 5 independent experiments, +/- SEM.