DNA binding of mutant and wild-type FANCJ proteins as detected by gel mobility shift assays. (A) The indicated concentrations of FANCJ-WT or FANCJ-A349P protein were incubated with 0.5nM forked duplex DNA substrate at 25°C for 30 minutes, as described in supplemental Methods. The DNA-protein complexes were resolved on native 5% polyacrylamide gels. (B) Quantitative analyses of DNA gel-shift experiments as performed in (A). (C) Gel mobility shift experiments with radiolabeled single-stranded 67-mer oligonucleotide. (D) Quantitative analyses of DNA gel-shift experiments as performed in (C). (E) 2.4nM FANCJ-WT or FANCJ-A349P was incubated with 0.5nM radiolabeled forked duplex DNA at 25°C for 15 minutes, and the indicated concentration of 67-mer ssDNA was subsequently added and incubated for an additional 15 minutes at 25°C. DNA-protein complexes were resolved on native 5% polyacrylamide gels. Quantitative analyses of DNA-binding data from DNA competition experiments are shown with SD indicated by error bars.