Pd173074

The antibodies used are against N-cadherin, E-cadherin, plakoglobin (BD Biosciences; San Jose, California); fibronectin, cytokeratin 18, β-actin (Sigma; St. Louis, MO); Slug, vimentin, p-ERK, p-Akt, p-p53, Akt, Bcl-2, Bcl-xL, Bax, Bim, Puma, cleaved caspase-3 and PARP (Cell Signaling; Danvers, MA); Erk, Bax, Noxa and FGFR1 (Santa Cruz; Santa Cruz, CA). Drugs used are PD173074 and PD0325901 (Pfizer; Groton, CT), Iressa or ZD1839 (AstraZeneca; Wilmington, DE), MK2206 (Tocris; Bristol, United Kingdom).

Human MM cells were cultured in IMDM (Gibco,), while HEK293 cells were maintained in DMEM (Gibco,) and both media types were supplemented with 10 % FBS (Hyclone Laboratories, Thermo Scientific, South Logan) and penicillin (10,000 U/ml)/streptomycin (10,000 mg/ml; Bioshop, Burlington). All cells were maintained in 5 % CO₂ incubators at 37 °C. PD173074 (Pfizer, Ann Arbor, MI) treatment was performed at a final concentration of 100 nM for 4 h, essentially as described previously [3 (link)]. FGF1 (Cedarlane, R&D Systems) was used at 80 ng/ml with sodium heparin salt (30 μg/ml; Sigma) to activate FGFR3. Pervanadate (50 μM) was generated by oxidation of sodium orthovanadate (Calbiochem) with H₂O₂. Tunicamycin (Sigma-Aldrich) was used for 16 h at 1 μg/ml.