Figure 6

Electrophoretic mobility-shift assay employing cell nuclear extract from MCF7. (a) Gel shift analyses were performed with double-stranded oligonucleotides (27 bp) as indicated: positive control probe containing the GATA3-binding element (GATA3-wt), a mimetic wild-type MUC1 probe containing the predictive GATA3-binding site (MUC1-wt), and a probe sequence with predictive GATA3-binding site mutations (MUC1-mut). (b) The presence of GATA3 was confirmed by supershift assay. Nuclear extract prepared from MCF7 cells was preincubated with GATA3 antibody, and then incubated with the mimetic wild-type MUC1 probe containing the GATA3-binding site (MUC1-wt). Arrows to the right indicate the position of the free probe (FP), the probe–nuclear-protein complex (Complex), and the specifically retarded protein–GATA3-antibody–probe complex (Supershift).