Fig 1: Analysis of matrix metalloproteinase-9 (MMP-9) expression and inflammatory cell recruitment and collagen metabolism in thrombus resolution. (A and B) Expression level (A) and activity (B) of MMP-9 were analyzed in vein endothelial cells in rats with heparin-induced deep venous thrombosis. (C) The number of macrophages in vein endothelial cells after treatment with rivaroxaban. (D) Expression of von Willebrand factor (vWF) in endothelial cells from rivaroxaban-treated rats with deep venous thrombosis. (E) Analysis of the effects of MMP-9 on collagen metabolism in vein endothelial cells. (F) Viability of vein endothelial cells after treatment with rivaroxaban. (G and H) Improvement in elastin fibers (G) and the stiffness of collagen (H) were analyzed after thrombus resolution by rivaroxaban. All data are represented as means ± SEM of triplicate samples. One-way ANOVA revealed a significant effect. **P<0.01 vs. the control.
Fig 2: A20 affects MMP-9 and ZO-1 and maintains the integrity of the BBB after SAH. (A, B) Lentivirus transfection for A20 expression affected the expression of MMP-9 and ZO-1 as revealed by representative western blots. (C) A20 expression affected the activity of MMP-9 as revealed by ELISA. (D) Evaluation of the integrity of the BBB by injection of Evans blue dye. A20 played a role in maintaining the integrity of the BBB after SAH. Data are expressed as mean ± SEM, n = 6, *p < 0.05, **p < 0.01, ***p < 0.005, ns versus SAH. ns, non-significant.
Fig 3: IL-1β induced gene expression of gelatinases and TIMPs in MAPK-specific inhibitor-pretreated mono-cultured and co-cultured chondrocytes. (a) IL-1β at different concentrations induces gene expression of gelatinases and TIMPs in mono-cultured and co-cultured chondrocytes.1, 5, 10 and 20 = 1, 5, 10 and 20 ng·mL−1. GAPDH and β-actin were used as the reference genes. The product sizes are indicated in the left lane. The gels shown are representative of three independent experiments (n = 3). (b) High ratios of MMP-2/TIMP-2 and MMP-9/TIMP-1 induced by different concentrations of IL-1β in inhibitor-pretreated mono-culture and co-culture chondrocytes. The data were the mean of three different experiments (n = 3). *Significant difference with respect to the normal ratios (P < 0.05). (c) The gene profiles of the gelatinases and TIMPs induced by IL-1β and/or specific inhibitors in mono-cultured and co-cultured chondrocytes. GAPDH and β-actin were used as internal controls. The product sizes are indicated in the left lane. The gels shown are representative of three independent experiments (n = 3). (d) Different ratios of MMP-2/TIMP-2 and MMP-9/TIMP-1 inhibitor-pretreated mono-cultured and media co-cultured chondrocytes. The data are the mean of three different experiments (n = 3). *Significant difference with respect to control ratios (P < 0.05). (e) qPCR was done to confirm the gene expressions of gelatinases induced by IL-1β in mono-culture and co-culture group. *Significant difference with respect to monolayer chondrocytes (P < 0.05). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IL, interleukin; MMP, metalloproteinase; TIMP, tissue inhibitors of metalloproteinase.
Fig 4: IL-1β increases the activity of gelatinases in a dose-dependent manner in both mono-cultured and co-cultured chondrocytes. (a) Zymography showed different dose-dependent increases of the gelatinases in mono-cultured and co-cultured chondrocytes. Pro-MMP-9 (87 kDa), active-MMP-9 (83 kDa), pro-MMP-2 (68 kDa) and active-MMP-2 (65 kDa) are in the right lane. The gels shown are representative of three different experiments (n = 3). (b) The quantification was performed with Quantity One 4.6.3 software. The optical densities of the pro- and active-MMP-2 and -9 bands were added as the total value of activity for MMP-2 and -9. The data are the mean of three different experiments (n = 3). *Significant difference with respect to control (P < 0.05). (c) ELISA Kit confirmed the IL-1β-induced gelatinases secreted by chondrocytes in mono-culture and co-culture groups (mean ± standard deviation) (n = 4). *P < 0.05, **P < 0.025, compared with controls. ELISA, enzyme linked immunosorbent assay; IL, interleukin; MMP, metalloproteinase.
Supplier Page from Abcam for Mouse proMMP9 ELISA Kit