EMSA of HepG2 nuclear extracts.

HepG2 cells were treated for 4 hours with DFX (100 μmol/L) or CPX (8 μmol/L) under normoxic (20% oxygen) or hypoxic (1% oxygen) conditions. Nuclear extracts were incubated with a radioactively labeled oligonucleotide probe derived from the Epo 3′ HRE13 and separated by native polyacrylamide gel electrophoresis. Specific HIF-1 DNA binding was confirmed by supershift analysis using the monoclonal antibody mgc3.14