Figure 5. Identification of Ets-1 transcription factor as a protein interacting with −283/−252 oligonucleotide. A.

) Radiolabeled DNA probe encoding putative binding sites for Ets family of transcription factors (−283/−252) was incubated with 10–15μg of Hepa 1–6 and AML-12 nuclear cell lysate. The probe yielded similar band pattern with both the cell lysates. B) Competition EMSA was performed using labeled −283/−252 probe, Hepa 1–6 lysate and various competitor oligonucleotides at 50- and 200-fold molar excess. Competitor oligonucleotides used in EMSA are indicated above the figure. Specific bands are shown with arrows. C) Radiolabeled −283/−252 oligonucleotide was incubated with Hepa 1–6 nuclear lysate in the presence of either anti-PU.1 or anti-Ets-1 antibodies. DNA-protein complexes were resolved on 5% polyacrylamide gel and supershifted bands are shown with an open arrow. NE =  nuclear extract.