Restriction endonuclease accessibility of the erythropoietin promoter and 5′-UTR.

Hypoxic Hep3B and HeLa cells were incubated in vivo with Msp1 after a brief treatment with lysolecithin. DNA was purified and digested with Xba1 to create an amplification standard. Restriction enzyme cleavage of the DNA was detected by ligation-mediated polymerase chain reaction. X, nonspecific band. (arrow and bracket) Sites with increased resistance to Msp1 cleavage in HeLa. (filled circle) Xba1 site. The schematic diagram shows (dumbbells) Msp1 sites, (bent arrow) transcription start site, and (rectangle) GATA box. The ramp corresponds to increasing Msp1 concentration.