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100 nM Smad1 MH1 domain was bound to 1 nM 5′-Cy5-labelled 15-mer SBE DNA (lane 1). The bound protein–DNA complex was incubated with 2 µM unlabelled competitor DNA: 14-mer blunt SBE, 15-mer blunt SBE (positive control for complete competition), 16-mer blunt SBE, 17-mer TTAA overhang SBE and 16-mer SBE with spacer (lanes 2–6). An oligonucleotide with the mutated SBE sequence TCATCTGATTTATACT was used as a negative control (lane 7).
