Direct binding of PU.1 to Ets-motifs in the human pIII promoter.

A. Nucleotide sequences of probes and competitive oligonucleotides used for EMSAs. B and C. EMSA profiles. Anti-PU.1 Ab (P) or control Ab (C) was used to identify the specific band composed of PU.1 and probe DNA (B). EMSA with an excess amount of competitors (a 1- or 10-fold molar concentration of probe 1 or 2) was performed to identify PU.1-binding sites (C). λB, which is previously published element bound with PU.1 [23], was mixed with a 15- or 30-fold concentration of competitors. Specific bands corresponding to complexes of PU.1 and the probe are marked with an asterisk. Super-shift band corresponding to complexes of PU.1, the probe, and anti-PU.1 Ab are marked with double asterisks. n.s., non-specific bands detected in protein mixture without probe DNA.