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EMSA analysis of NFAT DNA-binding activity in vitro. (A) The effective Kd of wild-type NFAT2-RHR for ARRE2 is estimated at 20 nM. (B and C) The K538A and R540 NFAT2 mutants bind ARRE2 with substantially lower affinity. From the data in A–C, the effective Kd of the wild-type, K538A and T551A mutants for ARRE2 is ≈20 nM, ≈165 nM, and ≈170 nM, respectively. Nonspecific binding of proteins to DNA was observed at high protein concentrations (lane 5 and 6 of A).
