FIGURE 2.

A, schematic representation of the HIV-1 subgenomic plasmid pDP, which is derived from pNL43 (48) and contains a deletion between nucleotides 1571 and 3516 (numbers refer to HXB2R sequence). The three HIV-1 mRNA size classes, and vpr mRNA 13a7, are indicated below plasmid pDP. Arrows indicate oligonucleotides used for RT-PCR. The Northern blot probe is indicated. B, RT-PCR on cytoplasmic RNA extracted from HeLa cells transfected with pDP in the absence or presence of pCMVSRp40 or pCMVSRp55. Duplicate transfections are shown. The RT-PCR was performed with the oligonucleotides Exon3s and BAMA (A). The different bands represent the indicated mRNAs. C, Northern blot of cytoplasmic RNA extracted from HeLa cells transfected with pDP in the absence or presence of increasing concentrations of pCMVSRp55 or pCMVSRp75. The mRNA size classes are indicated. *, the shift in migration of 2-kb mRNAs. D, the RNAs in C were used for RT-PCR with oligonucleotides Exon3s and BAMA (A) that detect mRNAs containing exon 3. E, RT-PCR on cytoplasmic RNA extracted from HeLa cells transfected with pDP in the absence or presence of pCMVSRp40, pCMVSRp55, or pCMVSRp75. The RT-PCR was performed with the oligonucleotides BSS and BAMA (A) detecting all HIV-1 mRNAs. The bands representing mRNAs are indicated. M represents the molecular weight marker.