Fig. 3.
EMSA with recombinant Gata-1 proteins on the inverted noncanonical double site.
(A) EMSA with a recombinant Gata-1 (rGata-1) molecule containing both C- and N-terminal zinc fingers (double-finger) binds FT1 probe containing both noncanonical Gata-1 sites, but not probes with the 5′, 3′, or both sites mutated (FT1B, FT1C, and FT1D, respectively). (B) EMSA performed with a recombinant Gata-1 protein containing only the C-terminal zinc finger (C-finger) binds all 3 probes that contain at least one noncanonical site (FT1, FT1B, and FT1C). Each probe was assayed with 10 mL 12.5, 50, and 100 nM DNA concentration.