Figure 1

BI-2 blocks the formation of 2-LTR circles during HIV-1 infection. Cf2Th cells were challenged with HIV-1 expressing GFP as a reporter (HIV-1-GFP) in the presence of increasing concentrations of BI-2 or PF74. Infection was determined 48 hours post-infection by measuring the percentage of GFP-positive cells by flow cytometry (A). Similar results were obtained in three independent experiments and a representative experiment is shown. Similarly, Cf2Th cells treated with BI-2, PF74 or DMSO were challenged with DNAse-pretreated HIV-1-GFP viruses. Subsequently, cells were harvested 7, 24 and 48 hours post-infection to measure HIV-1 late reverse transcripts (LRT) (B), formation of HIV-1 2-LTR circles (C) and infectivity (D), respectively. As control, we performed similar measurements in the presence of the reverse transcriptase inhibitor nevirapine (Nev) (B-D). Similar results were obtained in three independent experiments and standard deviations are shown. (E) Similarly, Cf2Th cells treated with the indicated concentrations of BI-2 were challenged with DNAse-pretreated HIV-1-GFP viruses. Subsequently, cells were harvested 7 and 48 hours post-infection to measure HIV-1 LRT (left panel) and infection (right panel), respectively. As a control, we performed similar experiments in the presence of the reverse transcriptase inhibitor nevirapine (Nev). Similar results were obtained in three independent experiments and standard deviations are shown. (F) Formation of Late reverse transcripts by HIV-1-GFP and HIV-1-T107N-GFP were measured at 7, 24 and 48 hours post-infection in the presence of the indicated amounts of BI-2 or PF-74. Viral Infection was determined by counting the percentage of GFP-positive cells 48 hours post-infection. Similar results were obtained in three independent experiments and standard deviations are shown.