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. 2002 Sep 15;30(18):4075–4087. doi: 10.1093/nar/gkf529

Figure 6.

Figure 6

Effect of plasmid conformation on probe binding to plasmid DNA in the presence of purified DNA-PKcs/Ku. Mixtures of pUC18 and pMII-MAR at 0.5 µg each were preincubated without or with 0.5 µg of M059K nuclear extract for 10 min at room temperature, purified using phenol/chloroform and ethanol precipitated. Agarose mobility shift assays containing labeled probe (1 ng) and untreated or nuclear extract pretreated plasmid DNA were performed with increasing amounts of purified DNA-PKcs/Ku as indicated. A separate reaction containing labeled probe, M059J nuclear extract (0.5 µg) and 30 ng of DNA-PKcs/Ku [(A), lane 10] was included as a positive control. Reaction mixtures were separated in a 1.5% agarose gel, dried, and subjected to autoradiography (A). A duplicate set of reactions was separated in parallel but stained with 1 µg/ml ethidium bromide and photographed (B).