Figure 2 of Andreae, Mol Vis 2020; 26:540-562.


Figure 2. The proteolytic activity of thrombin is required for increasing the Cyr61 protein synthesis in human corneal stromal fibroblasts and myofibroblasts. A and B: Fibroblasts (A) and myofibroblasts (B) were treated with a serum-free medium alone (-), a medium with 1.0 U/ml thrombin (+), a medium with 4.4 AT-U/ml hirudin, or a medium with 1.0 U/ml thrombin + 4.4 AT-U/ml hirudin. Cell lysates were collected, processed, and evaluated via western blot analysis for Cyr61 using the central linker-specific Cyr61 antibody. Representative blots are shown. GAPDH was used for normalization. Intracellular band densities from four independent experiments using different cell donors were analyzed via the two-way ANOVA. For fibroblasts, Hirudin Inhibitor factor: F: 10.973, df: 1, 12, p = 0.006; Thrombin factor: F: 6.527, df: 1, 12, p = 0.025; Interaction factor: F: 1.423, df: 1, 12, p = 0.256; n = 4. For myofibroblasts, Hirudin Inhibitor factor: F: 12.536, df: 1, 12, p = 0.004, n = 4; Thrombin factor: F: 8.116, df: 1, 12, p = 0.015, n = 4; Interaction factor: F: 3.816, df: 1, 12, p = 0.074; n = 4. Statistically significant comparisons and corresponding p values for the Tukey multiple comparison tests are denoted in the figure as *p = 0.002, **p = 0.005, ***p = 0.008, ****p = 0.021, given above the bars. Error bars displayed are SEM.