Mfm223 cells

CAL120 cells were a gift from Elgene Lim (Garvan Institute of Medical Research, Darlinghurst, NSW 2010, Australia). MFM-223 cells were purchased from Sigma-Aldrich. MDA-MB-453 cells were purchased from ATCC (Manassas, VA, USA). SUM185PE cells were purchased from Asterand Bioscience. Cells were cultured in RPMI-1640 (Gibco) and supplemented with 10% (v/v) FBS (Moregate), 10 μg/ml Actrapid penfill insulin (Clifford Hallam Healthcare) and 20 mM HEPES (Gibco).
For harvesting, cells at 80% confluency were washed twice with ice cold 1 × PBS then lysed with RIPA buffer (0.5% (w/v) sodium deoxycholate, 150 mM NaCl, 1% (v/v) NP40, 50 mM Tris–HCl pH 8.0, 0.1% (w/v) SDS, 10% (v/v) glycerol, 5 mM EDTA and 20 mM NaF), supplemented with 10 μg/ml aprotinin, 1 mM PMSF, 10 μg/ml leupeptin, 1 mM sodium orthovanadate, 2.5 mM sodium pyrophosphate and 2.5 mM β-glycerophosphate prior to use. Lysed cells were collected and clarified by centrifugation at 21,130 × g at 4 °C for 10 min, then the protein concentration was determined using a Pierce BCA protein assay kit (Thermo Scientific) according to the manufacturer’s protocol.

MCF-7 cells were purchased from the American Type Culture Collection (ATCC) and MFM223 cells from Sigma-Aldrich. MDA-MB-231(SA) cells were a gift from Dr. Guise (University of Texas, USA). MDA-MB-231(SA) is a variant of MDA-MB-231 that effectively forms bone metastases, but does not otherwise markedly differ from the parental cell line [22 (link), 23 (link)]. MCF-7 cells were grown in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% heat-inactivated fetal bovine serum (iFBS, Gibco), GlutaMAX (Gibco), estrogen (10^–9 M) and insulin (4 μg/ml). MDA-MB-231(SA) cells were grown in 10% iFBS/DMEM (Sigma-Aldrich) supplemented with GlutaMAX and non-essential amino acids (Gibco). MFM223 cells were grown in 10% iFBS/DMEM. The FGFRis TKI258 (S2769), BGJ398 (S2183) and AZD4547 (S2801) were purchased from Selleck Chemicals as DMSO stocks (IC50 values are listed in Supplementary Table S1).

The BT549, BT20, DU4475, HCC38, HCC70, HCC1500, HCC1569, HCC1954, HCC1806, HCC1143, HCC1937, HS578T, MDA-MB-157, MDA-MB-436, MDA-MB-453, MDA-MB-231, and MDA-MB-468 cell lines were purchased from the American Type Culture Collection; CAL51, CAL148, and CAL851 cells were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen; SUM185PE and SUM149PT cells were purchased from Asterand Bioscience; MFM223 cells were purchased from Sigma-Aldrich; and CAL120 cells were a gift from Professor Elgene Lim from the Garvan Institute of Medical Research, Darlinghurst, NSW, Australia. All the above cell lines were cultured in RPMI supplemented with 10% FBS and 0.023 IU/ml (or 10 µg/ml) insulin. MDA-MB231(LM2) cells (a kind gift from Dr. Joan Massague, Sloan-Kettering Memorial Institute, New York, NY; Minn et al., 2005 (link)) were cultured in DMEM supplemented with 10% FBS.