Figure 4

PHRF1 modulates NHEJ. HindIII-linearized pGL3 plasmid containing the luciferase gene was transfected into PHRF1-depleted HEK293T cells (a) and PHRF1-overexpressing U2OS cells (b), respectively. In vivo end-joining efficiency was measured by luciferase activity and normalized with β-gal expression. The data shown were mean±S.D. of three independent experiments. NHEJ reporter H1299dA3-1 no. 2 cells were transfected with PHRF1 siRNAs (c) or HA-PHRF1 (d) for 24 h and then transfected with I-SceI expression plasmid for another 48 h. The proportion of enhanced green fluorescent protein (EGFP)-positive cells was determined by flow cytometry at 48 h after I-SceI transfection. Each experiment represents the mean±S.D. of three independent experiments. *P<0.05 and **P<0.01 compared with the control. HA, hemagglutinin; IP, immunoprecipitation