Fedratinib

Ruxolitinib (CAS 941678–49-5, Cat#11609) and fedratinib (TG101348) were purchased from Cayman Chemical, dissolved in DMSO, and stored as 5 μl stock aliquots: 20 mM Ruxolitinib at −20 °C and 10 mM fedratinib at −80 °C. Recombinant human TNF-α was obtained from R&D Systems (#210-TA). Nuclear extracts were prepared by using the kit from Cayman Chemical (#10009277) following the manufacturer’s guidelines. Protein was quantified by using the Pierce BCA Protein Assay Kit (ThermoScientific Cat#23225). Human factor X (#HCX-0050) and human factor VIIa (HCVIIA-0031) were purchased from Haematologic Technologies. Chromogenic substrate for the determination of factor Xa activity (Chromogenix S-2765, Diapharma) was reconstituted per manufacturer’s instructions and stored at 4 °C. The urokinase type plasminogen activator human chromogenic activity assay kit was purchased from Abcam (Cat#ab8915) and used following the manufacturer’s protocol. HEPES buffered saline [HBS] (20 mM HEPES, pH 7.4, 150 mM NaCl) was used for tissue factor assay with the addition of 3% w/v bovine serum albumin (Fraction V; Fischer). CaCl2 (10 mM, final concentration) was added. For quenching the reactions, EDTA (10 mM final concentration) was added.