Incb18424

Human ovarian cancer cell lines (SKOV-3, 2774 and OVCAR-3) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Human IFN-α and IFN-γ were obtained from Sigma (St. Louis, MO). The chemical doxazosin was also purchased from Sigma. The JAK1/2 kinase inhibitor INCB18424 (Ruxolitinib) and STAT1 inhibitor (NSC118218) were obtained from Selleck Chemicals (Houston, TX), and stock solutions were prepared in DMSO. NSC74859 (S31-201), a specific STAT3 inhibitor, was purchased from Calbiochem Chemicals (La Jolla, CA). The following primary antibodies were used in this study: anti-JAK1, anti-phospho-JAK1, anti-JAK2, anti-phospho-JAK2, anti-STAT1, anti-phospho-STAT1, anti-STAT3, anti-phospho-STAT3, anti-caspase-3, anti-cMyc, anti-Bcl-2, anti-Bax, anti-p53, anti-survivin, and anti-COX-2 (Cell Signaling, Beverly, MA), anti-PARP, anti-XIAP (BD Biosciences, San Jose, CA), anti-cyclin D1, anti-CDK4, anti-Akt, anti-phospho-Akt, anti-TYK2, anti-phospho-TYK2, anti-PI3K, anti-phospho-PI3K, anti-mTOR, anti-phospho-mTOR, anti-PKCδ, anti-phospho-PKCδ, anti-STAT2, anti-phospho-STAT2, anti-p70S6K, and anti-phospho-p70S6K (Santa Cruz Biotechnology, Santa Cruz, CA), anti-p21, and anti-p27 (Oncogene, San Diego, CA).

Human breast cancer cell lines (MDA-MB-468, MDA-MB-231 and MCF-7), lung cancer cell lines (A549 and H460), and colon cancer cells (HCT-116 and HT-29) were obtained from ATCC (the American Type Culture Collection, Manassas, VA, USA). Cells were grown in Dulbecco's modified Eagle's medium (DMEM) or RPMI 1640, containing 10% heat-inactivated fetal bovine serum. Alisertib (MLN8237) and INCB-18424 (Ruxolitinib) were purchased from Selleck chemicals (Houston, TX). Nocodazole and thiazolyl blue tetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO). VX-680 (Tozasertib) was purchased from BioVision incorporated (Milpitas, CA). AJI-214 and AJI-100 were synthesized as described previously [30 (link)]. All drugs for cell culture were dissolved in DMSO (Sigma).