Fig. 7.
Envelope incorporation in SHIVKU-1bMC33 and SHIVSCVpu. C8166 cells were inoculated with either SHIVKU-1bMC33 or SHIVSCVpu for 6 days. At day 6, cultures were labeled as described in the Materials and methods section. The virus culture supernatants were centrifuged through a 20% sucrose cushion and lysed for immunoprecipitation. SHIV proteins were immunoprecipitated from cell lysates using plasma pooled from several pig-tailed monkeys infected previously with SHIV as described in the Materials and methods section. Uninfected C8166 cells served as a negative control. All immunoprecipitates were collected on protein A Sepharose, the beads were washed three times with RIPA buffer, and the samples were resuspended in sample reducing buffer. Samples were boiled and the SHIV specific proteins analyzed by SDS-PAGE (10% gel). (Lane 1) Immunoprecipitated SHIV proteins from SHIVSCVpu.(Lane 2) Immunoprecipitated SHIV proteins from uninfected C8166 cells. (Lane 3) Immunoprecipitated SHIV proteins from SHIVKU-1bMC33. Molecular weight markers are to the right of the gel.