Figure 7: UVRAG^FS inhibits NHEJ repair.

(a) Neutral comet assay shows a delay of DNA DSBs repair in UVRAG^FS cells. SW480 cells stably expressing empty vector (first row), UVRAG^WT (second row) or UVRAG^FS (third row) were treated with 1 Gy IR. The DNA damage levels of the cells before IR, 10 min post IR and 24 h post IR were assessed. Representative comet images are shown in the left panel and quantifications are shown on the right. (b) UVRAG^FS inhibits UVRAG interaction with Ku70 and Ku80 and the interaction of Ku70/80 with DNA–PKcs. The 293T cells transfected with increasing amounts of Flag-UVRAG^FS were treated with IR (5 Gy). WCL were immunoprecipitated with anti-DNA–PKcs or anti-UVRAG, followed by immunoblotting with the indicated antibodies. (c) HEK293 cells stably expressing the EJ5-GFP reporter for NHEJ and the DR-GFP reporter for homologous recombination (HR) were transfected with an empty vector or Flag-UVRAG^FS before the induction of DSBs by SceI transfection. The DNA repair activities as assessed by the reconstituted GFP signals were quantified by fluorescence-activated cell sorting. Data shown represent mean±s.d. (n=3). **P<0.01.