Fig. 5

Impact of UNG2 and RPA32 on HIV-1 replication in PBMCs. a–c Viral replication. Replication-competent viruses were produced in shLuc- (black curves and bars), shUNG2- (red curves and bars) or shRPA32- (green curves and bars) transduced 293T cells, normalized for p24, and then used for infection in duplicate of PBMCs from five different healthy blood donors. Aliquots of PBMC culture supernatant were collected 2, 4 and 8 days after infection for p24 quantification. In a, the individual kinetics of replication in PBMCs from the five healthy donors are shown. In b, results are expressed as the percentage of p24 production at each time point relative to that of PBMCs infected with viruses produced in shLuc-transduced (black bars) 293T cells. Values are the means of two independent experiments performed on PBMCs from the five healthy donors. In c, PBMCs were collected 7 h after infection, subjected to DNA purification, and total viral DNA was quantified via qPCR using specific primers for U5-gag. Results are expressed as the percentage of total viral DNA relative to that of PBMCs infected with viruses produced in shLuc-transduced (black bar) cells. d Virus infectivity. GFP reporter viruses were produced in shUNG2-, shRPA32- or shLuc-transduced 293T cells as indicated, normalized for p24, and then used to infect PBMCs from three different donors. The percentage of GFP-positive infected cells was then measured by flow cytometry 60 h later. Viral infectivity was normalized to that of viruses produced in shLuc-transduced 293T cells. Error bars represent the SEM. Statistical significance was determined by using the Students t test (ns, p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001)