Figure 5.

Smooth muscle cell index and vascular endothelial growth factor receptor 1 (VEGFR-1) expression are reduced in lentivirus (LV)–small hairpin RNA (shRNA)–vascular endothelial growth factor-A (VEGF-A)–transduced vessels. (a, upper panel) Representative sections after α-smooth muscle actin (α-SMA) staining at the venous stenosis of the LV-shRNA-VEGF-A (LV) and scrambled shRNA (control (C)) and Avastin treated with control vessels at days 14 and 28. Cells staining brown are positive for α-SMA. immunoglobulin G (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 and C groups and Avastin-treated and control vessels are shown in a (lower panel). This demonstrates a significant reduction in the average α-SMA index in LV-transduced vessels when compared with C vessels by day 21 (P<0.001) and day 28 (P<0.01). There is also a significant decrease in the average α-SMA index in Avastin-treated vessels when compared with controls by day 14 (P<0.001). (c) Pooled data from RT-PCR analysis for VEGFR-1 expression after transduction from the LV and C groups. A typical blot is shown in the upper panel and the pooled data in the lower panel (c). This demonstrates a significant reduction in the mean VEGFR-1 expression in the LV-transduced vessels when compared with C vessels at day 7 (P<0.001). (b, c) 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<0.01, **P<0.001, or ^#P<0.0001.