Figure 6.

There is decreased hypoxia-inducible factor-1α (HIF-1α) expression and staining in lentivirus (LV)–small hairpin RNA (shRNA)–vascular endothelial growth factor-A (VEGF-A)–transduced and Avastin-treated vessels. (a) Real-time PCR (RT- PCR) analysis for HIF-1α expression after transduction with LV and control shRNA (C). A typical blot is shown in the upper panel and the pooled data in the lower panel. This demonstrates a significant reduction in average HIF-1α expression in the LV-transduced vessels when compared with C vessels at days 7 (D7; P<0.001) and 14 (D14; P<0.01). (b) Representative sections after HIF-1α staining at the venous stenosis of the LV-shRNA-VEGF-A (LV) and C and Avastin- and immunoglobulin G (IgG)–treated vessels at day 14 and day 28. Brown-staining cells are positive for HIF-1α. IgG antibody staining was performed to serve as negative control. *Indicates the lumen. All are original magnification × 40. Bar=200 μm. Pooled data for the LV-transduced and C vessels and Avastin-treated and control vessels are shown in b. This demonstrates a significant reduction in the average HIF-1α index in the LV-transduced vessels when compared with C vessels at days 14 (P<0.001) and 21 (P<0.01). There is a significant reduction in the average HIF-1α index in the LV-transduced vessels when compared with C vessels at days 14 (P<0.001) and 21 (P<0.01). There is no difference in the Avastin-treated vessels compared with controls at either time point. Each bar shows mean±s.e.m. of 4–6 animals per group. Two-way analysis of variance (ANOVA) followed by Student's t-test with post hoc Bonferroni's correction was performed. Significant difference from control value is indicated by ^#P<0.001.