Fig 2. Binding of nuclear proteins to ERE half-sites of the TFPI gene by EMSA.

MCF7 cells were treated ±10 nM EE2 for 4 h and nuclear proteins were isolated and incubated with biotin-labelled mutant or wild type oligonucleotides containing the ERE half-sites of interest as described in Table 1. (A) ERE half-site position -845, (B) ERE half-site position -769 and (C) ERE half-site position -50. The resulting complexes were resolved by non-denaturing polyacrylamide gel electrophoresis. For competition, a 200-fold excess of unlabelled specific (of corresponding sequence) or ERE consensus oligonucleotide were added during the pre-incubation period. The arrows indicate specific DNA-protein complex formation.