Fig. 3

HSV-1 gB, gD, and gH/gL do not significantly affect the synthesis and processing of HIV-1 viral proteins. 293 cells were transfected with either the empty pcDNA3.1(+) vector or one expressing HSV-1 gB, gD, or gH/gL and pNL4-3. At 30 h post-transfection, cells were starved for methionine/cysteine and radiolabeled with ³⁵S-methionine/cysteine for 16 h. The culture medium was collected and cell lysates prepared as described in the Materials and Methods section. HIV-1 proteins were immunoprecipitated using anti-HIV-1 antibodies while HSV-1 proteins were immunoprecipitated with gB, gD, gH, or myc (gL) antibodies. The immunoprecipitates were collected on protein-A-Sepharose, washed, and boiled in sample reducing buffer. The proteins were separated on 7.5% SDS-PAGE and visualized using standard radiographic techniques. a Immunoprecipitation of HIV-1 and gD proteins from cell lysates or culture medium from 293 cells co-transfected with either pcDNA3.1(+) and pNL4-3 or a vector expressing gD and pNL4-3. b Immunoprecipitation of HIV-1 and gB proteins from cell lysates or culture medium from 293 cells co-transfected with either pcDNA3.1(+) and pNL4-3 or a vector expressing gB and pNL4-3. c Immunoprecipitation of HIV-1 and gH/gL proteins from cell lysates or culture medium from 293 cells co-transfected with vectors pcDNA3.1(+) and pNL4-3 or vectors expressing gH/gL and pNL4-3. Shown below each cell lysate panel is the expression of β-actin as a loading control