Figure 5
Effect of oxygen normalization on expression of HIF-1α, HIF-2α, VEGF, and GLUT-1 protein expression in HUVECs and ASMCs. HUVECs (A-D) and human ASMCs (E-H) were exposed as described in Figure 1. Western blots were performed against HIF-1α (A and E), HIF-2α (B and F), GLUT-1 (D-H) on cellular lysates and ELISA was performed against VEGF (C and G) on cellular supernatants. Graphs indicate the densitometric analysis relative to the respective normoxic protein expression (dashed line) with a representative blot for each protein above each graph. Acute hypoxia (2.5% 02; 4 hours – "AH") was used as a positive control for the induction of each of the target protein. Data in graphs represent the mean +/- S.E.M. from three separate experiments with a total sample size of 4–6 and with representative gels above each graph. Asterisk, P < 0.05; double asterisk, P < 0.01; triple asterisk, P < 0.001 as compared to cells incubated in normoxia for three weeks.