Extended Data Fig. 9 |. UAF1 stimulates RAD51AP1-mediated R-loop formation in vitro.
a, In vitro D-loop formation with RAD51 or the RAD51AP1–UAF1 complex. RAD51 was incubated with labelled 60-nt ssDNA and then with RAD51AP1 or the RAD51AP1–UAF1 complex. A dsDNA plasmid containing a sequence homologous to the ssDNA was then added to the reactions. Formation of D-loops was analysed by native gel electrophoresis. Representative results from 2 similar experiments are shown. b, In vitro R-loop formation with the RAD51AP1–UAF1 complex. Preformed RAD51AP1–UAF1 complexes were incubated with labelled 63-nt ssRNA and then with a dsDNA plasmid containing a sequence homologous to the ssRNA. Formation of R-loops was analysed by native gel electrophoresis. The efficiency of R-loop formation was determined by quantifying the shifted and unshifted bands in a light exposure of the gel. Data are mean ± s.d. (n = 3 independent experiments).