FIG. 5.

Effects of PNA_PR2 on Gag-Pol expression and virion production in HIV-1-infected cells (A and B) and on Gag protein expression in HLtat cells transfected with the Gag expression plasmid p55M1-10 (C). (A) The amounts of viral RNA in the culture supernatant (pelleted virion derived) (lanes 1 to 3) and in the cells (lanes 4 to 6) were evaluated by RT-PCR (top). Arrows indicate the pertinent PCR products for each primer pair. The virion-derived Gag protein in the culture supernatant (lanes 1 to 3) and intracellular Gag protein processing (lanes 4 to 6) were evaluated by Western blot analysis using anti-p24^Gag antibody (bottom). Shown are the positions of Pr160^Gag-Pol, Pr55^Gag, p41, and p24 Gag proteins. Lanes of each gel represent H9_LAI cells cultured alone (lanes 1 and 4) or incubated with 100 μM PNA_PR2 (lanes 2 and 5) and 2 μM indinavir (lanes 3 and 6). (B) Western blot analyses with anti-p24^Gag antibody and anti-p17^Gag antibody (top) and with anti-p6^Gag antibody (bottom) of cell lysates processed from H9_LAI cells cultured alone (lane 1) or in the presence of 30 μM PNA_PR2 (lane 2), 60 μM PNA_PR2 (lane 3), 2 μM indinavir alone (lane 4), 30 μM PNA_PR2 plus 2 μM indinavir (lane 5), or 60 μM PNA_PR2 plus 2 μM indinavir (lane 6). Cell lysate of uninfected H9 cells is shown in lane 7. PI − or + represents the absence or presence of a protease inhibitor, indinavir, respectively. The level of p24 antigen in each culture supernatant is indicated at the bottom. (C) Western blot analyses with anti-p24^Gag antibody (left) and anti-p6^Gag antibody (right) of cell lysates processed from HLtat cells transfected with pHXB2-based plasmid pSUM9 (136) (lanes 1) or with p55M1-10 and cultured alone (lanes 2) or in the presence of 100 μM PNA_RR2 (lanes 3). The cell lysate of mock-transfected HLtat cells is included as a reference (lanes 4). Sup, supernatant; Ag, antigen; Ab, antibody.