Figure 8

CG-induced peritoneal angiogenesis and VEGF-A expression required CTGF. (a) The localization of CD31⁺ vessels in fibrotic peritoneum. Peritoneal sections at day 21 were obtained from mice treated with control IgG or FG-3019. Representative tissue sections stained with anti-CD31 antibody are shown. Bars, 50 μm. (b) Numbers of CD31⁺ vessels in the peritoneum are expressed as the mean number ± SEM per HPF (n = 5 mice/group). (c) Immunostainings of VEGF-A⁺ cells in fibrotic peritoneum at day 21. Representative tissue sections stained with anti-VEGF-A antibody are shown. Bars, 50 μm. (d) Numbers of VEGF-A⁺ vessels in the peritoneum are expressed as the mean number ± SEM per HPF (n = 5 mice/group). (e) Representative tissue sections of dual-immunostainings of GFP (green) and VEGF-A (red) at 21 days. Bars, 50 μm. (f) VEGF-A expression in PMCs stimulated with 5 ng/ml TGF-β₁. (g) VEGF-A expression in PMCs. PMCs were transfected with control siRNA or CTGF siRNA, and then stimulated with TGF-β₁ (5 ng/ml for 24 hrs). (h) VEGF-A expression in PMCs. PMCs were preincubated with FG-3019 (10 μg/ml) or control IgG (10 μg/ml) for 1 h, and then stimulated with TGF-β₁ (5 ng/ml for 24 hrs). (i) VEGF-A expression in NIH3T3 cells stimulated with 5 ng/ml TGF-β₁. (j) VEGF-A expression in NIH3T3 cells. NIH3T3 cells were transfected with control siRNA or CTGF siRNA, and then stimulated with TGF-β₁ (5 ng/ml for 24 hrs). (k) NIH3T3 cells were preincubated with FG-3019 (10 μg/ml) or control IgG (10 μg/ml) for 1 h, and then stimulated with TGF-β₁ (5 ng/ml for 24 hrs). In in vitro studies, all data are expressed as copies of VEGF-A mRNA relative to copies of β₂ microglobulin mRNA ± SEM (n = 3 cell preparations/group).