FIGURE 6.

WRN and Pol δ form a complex on X-junction DNA. WRN (∼50 fmol) and Pol δ (∼1000 fmol) were preincubated either separately or together on ice for 5 min. After the addition of DNA (100 fmol) and ATPγS, reaction mixtures were incubated at room temperature for 15 min. For supershift assays, anti-WRN antibody was added, and the reactions were incubated for an additional 15 min at room temperature. The reactions were transferred to ice for 15 min, and after the addition of glycerol to a final concentration of 15%, aliquots were electrophoresed through 3% non-denaturing gels at 290 V at 4 °C until the bromphenol blue front was 7 cm from the point of application. Gels were dried and developed on a PhosphorImager to visualize free and protein-bound DNA complexes. Complexes formed by Pol δ and WRN in the absence (A) or presence (B) of anti-WRN antibody are highlighted by dashed rectangles.