Electrophoretic mobility shift assays of RAD51 with the canonical nucleosome and taillessΔ19 H4 nucleosome. The binding reaction was performed in the presence of ADP, and complex formation was analysed by non-denaturing 4% polyacrylamide gel electrophoresis with ethidium bromide staining (left). The RAD51 ring (without linker DNA binding) bound to the RAD51 ring (with linker DNA binding)–nucleosome complex is separately detected as the band that migrates more slowly than the RAD51 ring–nucleosome complex. The binding ratios of the RAD51 ring (without linker DNA binding) to the RAD51–nucleosome complex, as illustrated in the red rectangle on the right side of the gel, were estimated. The average values of three independent experiments (shown in Supplementary Fig. 1a,b ) are plotted against the RAD51 concentration (right). Data are mean ± s.d. ( n = 3 independent replicates). P values were obtained by two-sided Welch’s t-test. Asterisks indicate significance at P < 0.05.
