Cgp57380

General tissue culture materials were obtained from VWR International. Antibodies against eIF4E, tubulin, HSP90, ZEB1 and Dicer were obtained from Santa Cruz, while antibodies against p-eIF4E, MNK1, p-MNK1 and Drosha were purchased from Cell Signaling. Anti-GAPDH antibody was from Millipore, anti-E-cadherin antibody was obtained from BD Bioscience, while anti-vimentin antibody was from Abcam. Secondary antibodies were purchased from Sigma. CGP57380 was obtained from Santa Cruz. siRNAs against MNK1 and MNK2 were purchased from Dharmacon, ZEB1 siRNA was obtained from Life Technologies, while eIF4E and hnRNPA1 siRNAs were from Santa Cruz. Aldefluor assay kit was purchased from Stemcell Technologies.

Established GBM cell lines (U87, LN229, and LN18) were grown in DMEM (Thermo Fisher) with 10% FBS (Thermo Fisher) and gentamycin (1 mg/mL). The identity of established cell lines was authenticated by short-tandem repeat (STR) analysis (Genetica DNA Laboratories) in August 2016. PDX cell lines were grown in serum-free Neurobasal medium (Thermo Fisher) with N-2 Supplement at a 1X final concentration (Thermo Fisher), EGF (50 ng/mL), bFGF (25 ng/mL), and penicillin/streptomycin (10 U/mL). Patient-derived MES (83Mes) and PN (AC17PN) GSC lines have been previously described (2 (link), 21 (link)). GSCs were grown in DMEM/F12 with EGF (20 ng/mL), bFGF (20 ng/mL), heparin (5 μg/mL), B27 (2%), and gentamycin (1 mg/mL) in nonadherent flasks. All cells were grown at 37 C in 5% CO₂. Pharmaceutical-grade ATO (Trisenox) used for in vitro and in vivo experiments was purchased from Teva Pharmaceuticals. The MNK inhibitor, CGP57380, was purchased from Santa Cruz Biotechnology. The arsenic–biotin conjugate (As-Biotin) was purchased from Toronto Research Chemicals and has been previously described (23 (link)).