Figure 7.
Retroviral cDNA synthesis. MDTF or D17 cells expressing the indicated TRIM5 orthologues were infected for 12 hours with HIV-1GFP at a MOI yielding about 20% infected cells for the control cells. In addition, infection of cells expressing TRIMCyp was carried out in the presence or absence of 5 μM cyclosporine, and infection of control cells was done in the presence or absence of the reverse transcriptase inhibitor nevirapine (80 μM). Top panel, total cellular DNAs were extracted and an aliquot of each DNA sample was subjected to a 30-cycle PCR amplification using ODNs annealing to GFP sequences. PCR products were separated on an agarose gel and revealed with ethidium bromide. Bottom panel, as above but HIV-1GFP-specific DNAs were quantitated by real-time PCR, using dilutions of a plasmid containing the GFP sequence as a standard.