Figure 2

Specific detection of the antisense transcript in latently infected cells and transfected 293T cells (A) RT-PCR analyses were performed on RNA samples from J1.1 cells using the 24-6 RT primer and PCR primer combinations 26-6/26-5 (lanes 1,2,4,6) and 26-6/25-3 (lanes 3,5,7) (expected size of 384 bp and 437 bp, respectively). Samples were tested for DNA contamination (lanes 2–3; no RT and no RT primer) and endogenous RT priming (lanes 4–5; RT with no added RT primer). Lane 1 represents PCR analysis with no added cDNA or RNA. Lanes 6 and 7 show the results of PCR using 2 primers combinations. (B) 293T cells were transfected with 5 μg of pNL4.3 or pNL4.3ΔNar1 proviral DNA. RT-PCR analysis of RNA samples from transfected 293 T cells and controls were performed as in A. M = Lambda EcoRI/HindIII DNA marker (the asterisk indicates the 564 bp band). The arrows on the right side of panel Apoints to the specific signal.