FIG. 8.
Measuring HIV-1 protein production in cells and supernatant in the presence of siRNAs. CA p24 or unprocessed p55gag was monitored by Western blotting for cells (C) and supernatants (A and B), at days 2 and 5, of 293T cells transfected with pNL4-3/env120-A or pNL4-3/env126-D and treated with siRNA120a or siRNA126a or mock treated. Both panels A and C employed a 1:400 dilution of patient sera derived from individuals infected with HIV-1 of either subtype A (JLT02A) or subtype C (JCR12) as a primary anti-p24 antibody, followed by peroxidase-conjugated secondary antibodies. (C) The anti-gp120 B13 antibody was employed in Western blot analyses with virus lysates obtained from day 5 supernatants of 293T cells transfected with pNL4-3/env and mock treated or treated with siRNA120a or siRNA126a. The B13 anti-gp120 antibody binds to a highly conserved linear epitope found in most HIV-1 gp120 sequences of most subtypes (panel B) (1)