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EMSA analysis of DNA-binding activity in vitro. (A and B) Wild-type AP-1 and R285A cJun mutant AP-1 bind with essentially identical affinities to a consensus AP-1 site (Kd = ≈80 nM) (A) and to the nonconsensus site in ARRE2 (Kd = ≈300 nM) (B). (C) The R285A cJun mutant AP-1 shows a substantially diminished ability to cooperate with NFAT1. From the data in C, the Kd of wild-type AP-1 for the NFAT1/ARRE2 complex is estimated at ≈30 nM.
