Figure 2

Western blot analysis of partially purified HIV-1 obtained from AdOx-treated cells. A) HIV-1 obtained from AdOx-treated or control CEM T-cells was pelleted through a 20% sucrose cushion. The pelleted virus was solubilized in RT lysis buffer and the CAp24 concentration was determined by ELISA. Western blot analysis was performed using 20 ng of CAp24 and probed with a human HIV-Ig. The proteins detected by the serum were visualized by ECL and two exposures at 2 minutes (left panel) and 20 seconds (right panel) are shown. The short exposure highlights that the amount of total virion protein was equal. The experiment was performed five times with similar results. B) HEK293T cells were treated with 20 μM AdOx 24 h. Equivalent amounts of HIV-1 obtained from cells was purified by centrifugation through 20% sucrose and resuspended in Berman Lysis buffer. After SDS-PAGE, western blot analysis was performed using a monoclonal CAp24 antibody. The results show one of three independent experiments that gave similar results. C) Whole cell lysates prepared from CHO cells or CHO cells stably expressing HIV-1 Env protein subunits gp120 and gp41 were analyzed by Western blot using a goat-anti HIV-1 polyclonal antibody (right panel) or HIV-Ig (left panel).