Figure 4. Incorporation of patient-derived Env glycoprotein into infectious virions.

293T cells were transfected with the indicated proviral expression constructs and the resulting chimeric viruses bearing patient-derived Env V1-V5 region were collected at 48hrs post-transfection. The viruses were pelleted through a 20% (wt/wt) sucrose cushion by ultracentrifugation from the clarified supernatants and the viral proteins were analyzed by Western blot. Gp41 protein was detected by anti-gp41 mAb 2F5, and p24 was detected by using anti-p24 mAb. Bands corresponding to gp41 and p24 are indicated. Env incorporation into virions was calculated as gp41/p24 ratio. For each pair, the NL4-3 value was set at 1, the relative Env incorporation for each virus was calculated relative to this value. UN means that the signal is undetectable.