Fig. 6

Both gD and gB are more efficiently released in presence of virus. In companion cultures from the experiment in Fig. 5, 293 were co-transfected with either 1) the empty pcDNA3.1(+) vector; 2) the vector expressing gD; 3) the vector expressing gD and pNL4-3; 4) the vector expressing gB; or 5) the vector expressing gB and pNL4-3. The cells were radiolabeled and harvested exactly as in Fig. 5. Following low speed centrifugation, the culture supernatants were layered onto a 20% sucrose cushion and virus pelleted by ultracentrifugation and different fractions were analyzed for the presence of gD or gB using immunoprecipitation. a Schematic showing the fractions analyzed: Fraction 1: medium prior to ultracentrifugation; Fraction 2: medium following ultracentrifugation; Fraction 3: the sucrose cushion following ultracentrifugation; and Fraction 4: virus pellet. b Immunoprecipitation of gD proteins from the different fractions of cells transfected with vector expressing gD. c Immunoprecipitation of gD proteins from the different fractions of cells co-transfected with the vector expressing gD and pNL4-3. d Immunoprecipitation of gB proteins from the different fractions of cells transfected with gB. e Immunoprecipitation of gB proteins from the different fractions of cells co-transfected with the vector expressing gB and pNL4-3. The amount of immunoprecipitated proteins run on gels represent 25% (Fraction 1), 25% (Fraction 2), 100% (Fraction 3) and 50% (Fraction 4) of the total immunoprecipitated product