Figure 5.

Mung Bean footprinting analysis of RPA and XPA binding to the damaged DNA bubble. The reaction mixtures (10 μl) contained 50 mM Tris–HCl 7.5, 100 mM KCl, 1 mM DTT, 0.6 mg/ml BSA, protein factors at analyzed concentrations and 10 nM 5′-³²P-labeled damaged DNA duplex. The left panel 5′-[³²P] labeled undamaged strand of DNA duplex. The right panel terminally labeled damaged strand of DNA duplex. After the Mung Bean DNA digestion reaction mixtures was separated on 10% denaturing polyacrylamide gel. The bottom panel shows schematic representation of the protection pattern for the DNA substrate. Strongly and weakly protected regions are designated by solid and shaded bars, respectively.