Identifier kit

The thyroid cell lines TPC1, Cal62, and 8505c were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). The sources of the cell lines have been described [57 (link)]. PCCl3-BRAF and PCCl3-HRAS were derived from PCCl3 rat thyroid follicular cells [58 (link)], to obtain doxycycline-inducible expression of BRAF^V600E or HRAS^V12 [57 (link), 59 (link), 60 (link)]. All cell lines used in this work were tested for mycoplasma contamination and authenticated every 6 months by short tandem repeat profiles using the Applied Biosystems Identifier kit in the Genomic Facility at the Institute of Biomedical Research (IIBm; Madrid, Spain).

The human HNSCC cell line CAL27 was purchased from the ATCC (Manassas, VA, USA). CAL27 is a naturally IL-6 low expressing cell line. IL-6 was overexpressed in CAL27 cells by using a retroviral vector as described previously [18 ]. UM-SCC-74A, a naturally IL-6 high expressing cell line, was obtained from Dr. Thomas E. Carey (University of Michigan). All tumor cell lines were cultured in DMEM supplemented with 10% fetal bovine serum, non-essential amino acids and penicillin/streptomycin. The identity of all cell lines was confirmed by STR genotyping (Identifier Kit, Applied Biosystems, Carlsband, CA). Bazedoxifene acetate was purchased from (Cayman Chemicals, Ann Arbor, MI) and cisplatin (CDDP) was purchased from Sigma-Aldrich (St. Louis, MO). Antibodies against pSTAT3, pERK1/2, pAkt, Survivin and XRCC-1 were obtained from Cell Signaling Technology (Danvers, MA). Nanog antibody for Western blotting was purchased from Novus Biologicals (Littleton, CO) and for immunohistochemistry was obtained from Abcam (Cambridge, CA). ERCC-1 antibody was purchased from Santa Cruz (Dallas, TX) and pFAK was from Invitrogen (Carlsband, CA). GAPDH antibody was obtained from EMD Millipore (Billerica, MA).