Asp3026 ct asp302

PPP was dissolved in DMSO for in vitro studies, and in DMSO/vegetable oil (10:1) for in vivo studies. ASP3026 (CT-ASP302; ChemieTek, Indianapolis, IN) was dissolved in DMSO with H₂O and HCl (1:1) for in vitro experiments, and in 0.5% methyl cellulose for in vivo experiments.

Five previously characterized NPM‐ALK⁺ T‐cell lymphoma cell lines: Karpas 299, SR‐786, SU‐DHL‐1, SUP‐M2, and DEL (DSMZ, Braunschweig, Germany), were used in the study (Drexler, 2010). The breast cancer cell line MCF7 and neuroblastoma cell line SK‐N‐AS (ATCC, Manassas, VA, USA) were used as positive controls for the expression of NGF and TrkA, respectively. Human peripheral blood CD3⁺ pan‐T lymphocytes were purchased from StemCell Technologies (catalog number: 70024; Vancouver, BC, Canada). The ALK inhibitor ASP3026 (CT‐ASP302; ChemieTek, Indianapolis, IN, USA) was dissolved in DMSO mixed with H₂O and HCl (1 : 1).
Cells were maintained in RPMI‐1640 medium (NPM‐ALK⁺ lymphoma cell lines) or Dulbecco's modified Eagle's medium (MCF7, SK‐N‐AS) (Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% heat‐inactivated fetal bovine serum (FBS), penicillin (100 U·mL⁻¹), streptomycin (100 μg·mL⁻¹), and l‐glutamine (2 mm) (all from Sigma Aldrich, St. Louis, MO, USA). Cell cultures were maintained at 37 °C in humidified air with 5% CO₂. In some experiments, cells were cultured overnight in 0.5–1% FBS. Then, 250 or 500 ng·mL⁻¹ recombinant human β‐NGF (256‐GF‐100/CF; R&D Systems, Minneapolis, MN, USA) was added with or without 5 μg·mL⁻¹ anti‐TrkA‐neutralizing antibody (AF175; R&D Systems).

ASP3026 (CT-ASP302; ChemieTek, Indianapolis, IN) was suspended in DMSO admixed with H₂O and HCl (1:1) for in vitro experiments and in 0.5% methylcellulose for in vivo experiments.