FIG. 2.
(A) Expression of HIV-1 tat, rev, and nef spliced mRNAs in H9 cells acutely infected with HIV-1IIIB. Lanes: 4 to 6, the HIV-1 tat (402 bp), rev (219/225 bp), and nef (203 bp) spliced mRNAs were expressed at 18 to 32 h postinfection; 8 to 12, β-globin mRNA expression from samples in lanes 2 to 6 (the densitometer reading of each band is shown under the gel); 7 and 13, RT-PCR DNA controls (RNA sample without the RT reaction). The aliquots of the cells in Fig. 1 were extracted for total cellular RNA, which was then digested with RNase-free DNase as previously described (36, 37). RT-PCR was performed as described in Materials and Methods. (B) Delayed expression of HIV-1 tat, rev, and nef spliced mRNAs in H9 cells acutely infected with HIV-1 NC mutants CCCC and SSHS. (Top) Lanes: 2 to 5, HIV-1 spliced mRNA was not detected in cells infected with NC mutant CCCC; 6 to 9, HIV-1 spliced mRNA of SSHS was expressed only at 48 h postinfection; 10 to 13, HIV-1 spliced mRNA of the wild type was expressed at 18 to 48 h postinfection. (Bottom) Lanes: 2 to 13, β-globin mRNA expression from samples in the top panel (the densitometer reading of each band is shown under the gel); 14, RT-PCR DNA control.