Ruxolitinib

SK-BR-3, A431, NIH-3T3 and A549 cell lines (American Type Culture Collection) were grown in DMEM or DMEM-F12 medium supplemented with 10% FCS at 37 °C in an humidified atmosphere of 5% CO₂. Cells were cultured up to passage 30. Thapsigargin (Sigma-Aldrich) is used at 1 μm. Lapatinib (2μM), CP-724714 (2μM), MK-2206 (1μM) and trametinib (10μM) were purchased from Selleckchem. LY294002 (25μM), gefitinib (1μM), and erlotinib (5μM) were provided by Tocris, ruxolitinib (1μM) by VWR and trastuzumab (1μM) by Roche.

The chemical inhibitors stattic, afatinib, and PD0325901, 3-Bromopyruvate, PFK-15, and Etomoxir were purchased from Selleck chemicals and used at indicated concentrations. Cytosine-1-β-D-arabinofuranoside (AraC) and cycloheximide were purchased from Sigma-Aldrich. Ruxolitinib was purchased from VWR. CPI-613 was purchased from Biovision Inc.
Antibodies against phospho-STAT3 (S727), phospho-STAT3 (Y705), and total STAT3 were purchased from Cell Signaling Technology. Anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH) antibody was purchased from Santa Cruz Biotechnology. Antibodies raised against VACV E3 protein were kind gift from Dr. Yan Xiang (UTHSA) [88 (link)]. Antibodies against VACV L2 protein were kindly provided by Dr. Bernard Moss (NIAID). A commercially synthesized recombinant VGF peptide corresponding to the cleaved fragment of VACV VGF [25 (link)] was purchased from GenScript.