Figure 2.

GSNO causes an accumulation of HIF-1α protein and HIF-1α DNA binding in normoxia, but not in the presence of PI3K inhibitors. (A) BPAECs or Cos 7 cells were treated with 100 μM GSNO for various times. HIF-1α protein was detected in whole cell lysate in treated and untreated cells by Western blot analysis using anti–HIF-1α antibody. Blot is representative of several. (B) Cos 7 cells were treated with 100 μM GSNO or hypoxia for the times indicated. HIF-1 binding to DNA was determined by electromobility shift assay on whole cell lysate. (C) Cos 7 cells and BPAECs were treated with 100 μM GSNO in the presence or absence of the PI3K inhibitors 100nM wortmannin or 25 μM Ly 294002. HIF-1α protein was detected in whole cell lysate by Western blot analysis using anti–HIF-1α antibody.