The cloned cDNA of the human homologue of EBF encodes a 57-kd protein that interacts specifically with a binding site in the human mb-1 promoter.
(A) Autoradiogram of 35S-labeled in vitro–translated mouse and human EBF. Positions of molecular-weight markers and EBF are indicated by arrows. (B) EMSA using the human mb-1 promoter EBF site in combination with either 0.5 or 2 μL of unprogrammed reticulocyte lysate (ret.) or 0.5 and 2 μL of recombinant in vitro–translated human EBF (hEBF). An unidentified DNA-binding activity present in the reticulocyte lysate is indicated by the star (*), and F indicates free probe. (C) EMSA in which competition was induced for the binding of in vitro–translated recombinant human EBF to the human mb-1 promoter EBF-binding site by the addition of duplex oligonucleotides spanning the wild-type (mb-1) and a point-mutated human mb-1 promoter EBF site (mut.mb-1), or the PyG box (PyG) from the human CD19 promoter, as indicated. The star indicates the background DNA-binding activity in the reticulocyte lysate, and F indicates free probe.
