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. Author manuscript; available in PMC: 2024 May 1.
Published in final edited form as: J Thromb Haemost. 2023 Feb 2;21(5):1366–1380. doi: 10.1016/j.jtha.2023.01.027

FIGURE 3.

FIGURE 3

Ruxolitinib and fedratinib decrease TNF-α mediated tissue factor and urokinase mRNA upregulation and decrease tissue factor and urokinase activity. HUVEC were treated with 0.4 μM ruxolitinib or 1 μM fedratinib JDB for 30 minutes followed by addition of 10 ng/mL TNF-α for 4 hours. (A) Tissue factor (Factor 3, F3) mRNA levels were normalized to control. (B) Plasminogen activator urokinase (PLAU) mRNA levels were normalized to control. For activity assays, HUVEC were treated with 0.4 μM ruxolitinib or 1 μM fedratinib for 30 minutes followed by addition of 10 ng/mL TNF-α for 4 hours JDB. (C). Measurement of tissue factor (TF)-dependent Xa generation was performed using Chromagenix Xa solution as described in methods. Results are normalized to untreated cells. (D). For urokinase activity, conditioned media were collected followed by urokinase activity measurement (Abcam). For all graphs, each dot represents an individual replicate from n = 2 to3 separate experiments. Error bars represent mean ±SE of means. p values from analysis of variance testing with appropriate post-hoc multiple comparison testing. ns, not significant. * p < .05, ** p < .01, *** p < .001, **** p < .0001.