Fig. 4.
Hypoxic-inducible HIF-1 DNA-binding activity in SH-SY5Y cells.
EMSAs of nuclear extracts prepared from SH-SY5Y cells exposed to anoxia for 4 hours were performed with the 32P-labeled EPOWt oligonucleotide. The HIF-1 complex, the constitutive (c), the unspecific band (u), and the free probe are indicated. Supershifted bands are indicated by arrows. Supershift analyses were performed with anti–HIF-1α (lane 1), anti-ARNT1 (lane 2), and anti-ARNT2 (lane 3) antibodies. Lane 4 represents the inducible HIF-1 band without any antibody. No inducible HIF-1 band was obtained when the nuclear extract was incubated with the EPOMut oligonucleotide (lane 5).
