Figure 2.

TFO AG30-mediated DNA repair synthesis of pUSAG15 and pUSAG16 in HeLa cell nuclear extracts. The supercoiled plasmid DNA was preincubated with oligonucleotide at 37°C for 2 h in triplex-binding buffer (10 mM Tris pH 7.5, 1 mM spermidine, 20 mM MgCl₂) for triplex formation. Then the plasmid DNA (1 µg) was incubated with HeLa nuclear extracts supplemented with dATP, dGTP, dTTP and [α-³²P]dCTP at 30°C for 2 h. The plasmid DNAs were linearized with XhoI restriction enzyme and analyzed by agarose gel electrophoresis using a 0.8% gel. (A) Visualization of the plasmid DNA by ethidium bromide staining. (B) Autoradiogram of the same gel showing labeled nucleotide incorporation indicative of DNA repair synthesis. (C) Quantification of incorporation of [α-³²P]dCTP into plasmid DNA. The amount of incorporated [α-³²P]dCTP in pUSAG15–AG30 was taken as 100%. Incorporation of [α-³²P]dCTP in other reactions was calculated as a percentage of pUSAG15–AG30. The data are means obtained from four individual experiments.