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

Figure 4.

Figure 4

RAD52 can compete with Ku for binding to HIV-1. (A) 293 cells transiently transfected with an increasing amount of HA-RAD52 expression plasmid, then infected with HIV-1 luciferase retroviral stocks. HIV-1 luciferase transduction assay results are shown (left) along with immunoblot analysis of cell lysates (right). Results are expressed as luciferase activity relative to untransfected 293 cells. Immunoblots were performed using anti-HA tag antibodies before re-probing for β-actin (loading control). (B) Competitive PCR–ChIP analysis of 293 cells transiently transfected with increasing amounts of HA-RAD52 expression plasmid and infected with HIV-1 luciferase retroviral stocks. Results show HIV-1 LTR PCRs for ChIPs using antibodies against the HA-tag, Ku80 and nonspecific controls (no antibody (−) and IgG1 isotype control anti-FLAG tag antibody). PCR amplifications were normalised to the amount of HIV-1 LTR DNA used per ChIP (INPUT DNA). (C) Immunoblot analysis of input extracts used to perform the ChIP assays. The amount of HA-RAD52 or Ku80 protein used per ChIP analysis was determined by immunoblotting using anti-HA tag or anti-Ku80 antibodies.