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. 2004 Aug 5;23(16):3421–3429. doi: 10.1038/sj.emboj.7600348

Figure 2.

Figure 2

Overexpression of RAD52 impairs retroviral integration. (A) Parental HeLa cells (HeLa) and clones stably transfected with vector DNA only (IRES-1 and -2) or stably overexpressing HA-RAD52 (RAD52-1 through -5) infected with HIV-1 luciferase retroviral stocks. HIV-1 luciferase transduction assay results are shown in the left graph and immunoblot analysis of the HeLa cell clones are on the right. Luciferase transduction assay results are expressed as luciferase activity relative to parental HeLa cells. Western blots of HeLa cell clones were performed using antibodies raised against the HA tag epitope, RAD52, Ku70, Ku80 and β-actin (loading control). (B) Quantitative real-time PCR analysis of late RT HIV-1 DNA isolated from Rad52−/− ES cells and HeLa cell clones overexpressing HA-RAD52 at 21 days after infection. The standard curve for quantification was generated from HeLa cell DNA isolated 24 h after infection (left panel). The amount of late RT HIV-1 DNA in each cell line (pg per 50 ng genomic DNA), determined from the standard curve, is shown in the right panel. UD: undetectable.