Figure 5. Effect of HIF-1α siRNA on hypoxia-induced VEGF expression. Bovine stromal cells were transfected with HIF-1α siRNA or non-targeting siRNA control for 6 h. Twenty-four hours after transfection cells were preconditioned with hypoxia for 4 h. A: Total RNA was collected immediately after treatment. Image shows RT-PCR analysis for HIF-1α and VEGF. 18s rRNA was detected as an internal control. B: Protein analysis of VEGF and HIF-1α. Whole cell lysates were collected immediately after treatment, separated by SDS PAGE and probed for VEGF and HIF-1α. β-actin was detected as a loading control. Bar graph shows VEGF expression in indicated groups relative to control. Error bars represent standard error of the mean (n=3).