Figure 8.
Regulation of inflammatory and fibrosis-related genes in vivo and of collagen production in vitro by TSP-4. A) Decreased levels of inflammatory markers in hearts of Thbs4−/− mice subjected to TAC. Real-time quantitative RT-PCR (qRT-PCR); levels of mRNA in TAC-operated WT mice = 100%; mean ± se, n = 5, 2 wk after TAC. Levels are normalized to corresponding values in sham-operated mice. B) Levels of MMPs in hearts of Thbs4−/− mice subjected to TAC. For real-time qRT-PCR, levels are normalized to corresponding values in sham-operated mice; means ± se, n = 5, 2 wk after TAC. C) Human foreskin fibroblasts (HFFs) were incubated with 50 μg/ml recombinant TSP-4 or gelatin for 24–48 h. Solubilized cells and their matrices were tested in dot-blot assays (n=3) using anti-collagen I-V, anti-collagen I, anti-collagen II, anti-collagen III, anti-collagen IV, anti-collagen V, or anti-β-actin (loading control) antibodies. Representative blot. D) Quantification of the results of experiment in C, n = 3. E) Cells [human umbilical vein endothelial cells (HUVECs), RF/6A microvascular endothelial cells, and HFFs] were treated as in C. Representative blot. F) Quantification of the results of experiment in E; n = 3.