FIG. 2.

Expression and incorporation of diverse HCV gp's into pseudotype particles. 293T cells were cotransfected with pNL4-3.Luc.R⁻E⁻ and plasmids expressing representative HCV gp's listed in Table 1 or an empty vector (No env). At 72 h posttransfection, the virus was pelleted through a sucrose cushion. The cells and virus were lysed, and both fractions were quantified for E2 expression by ELISA and Western blotting. The total E2 expressed per well of 293T cells (300 μg of cellular protein) (A) and that incorporated into pseudotype particles (B) (25 ng of particulate HIV p24 antigen) were measured by ELISA, and the data are expressed as optical density units (OD) at 450 nm and annotated above each bar. Virus preparations that gave optical density signals >2-fold the mean of the no-env virus control were considered to have incorporated gp's. Transfected cell lysate (10 μg of total protein) (C) and pelleted virus particles (D) (25 ng of particulate HIV p24 antigen) were separated by reducing SDS-PAGE and immunoblotted for E2 (MAb 3/11) and actin (MAb AC-15). The migration of molecular mass markers is indicated in kilodaltons.