Figure 3.

Western blotting of HIF-1α and the protein VEGF, encoded by a downstream target gene of HIF-1α. Myocardial cells were subjected to treatments simulating I/R (I/R), or they were mock-treated (Control). In parallel, cultures were transiently transfected with empty expression vector (I/R+NC) or an expression plasmid carrying full-length HIF-1α prior to I/R (I/R+pHIF). Total cell lysates were then prepared at 6 h and 5 days after I/R or mock treatment, and were subsequently analyzed using appropriate monoclonal antibodies, as detailed in the Materials and methods section. β-actin was used as a loading control. I/R, ischemia-reperfusion; HIF-1α, hypoxia-inducible factor-1α; VEGF, vascular endothelial growth factor.