Ba f3

The CML-derived cell line K562 and murine pro-B cell line Ba/F3 were purchased from the Riken Cell Bank (Ibaraki, Japan). K562 cells were cultured in RPMI 1640 (Nacalai Tesque) supplemented with 10% fetal bovine serum (FBS) (Biowest, Nuaillé, France), 100 units/mL penicillin (Nacalai Tesque), and 100 μg/mL streptomycin (Nacalai Tesque). Ba/F3 cells were cultured in RPMI 1640 supplemented with 10% FBS, 100 units/mL penicillin, 100 μg/mL streptomycin, and IL-3 (2 ng/mL).

The IL-3-dependent hematopoietic cell line Ba/F3 was purchased from Riken Cell Bank (Ibaraki, Japan). SUDHL-1 and Ki-JK cells, derived from NPM-ALK-positive ALCL patients, were purchased from Summit Pharmaceuticals International (Tokyo, Japan) and JCRB cell Bank (Osaka, Japan), respectively. SUDHL-1 cells and Ki-JK cells were cultured in RPMI 1640 medium supplemented with 10% FBS (BioWest), 100 units/ml penicillin (Nacalai Tesque), and 100 μg/ml streptomycin (Nacalai Tesque) with or without IL-3 (2 ng/mL) at 37°C and 5% CO₂. Ba/F3 cells were infected with an empty virus (-) and retroviruses expressing NPM-ALK, its kinase dead mutant (K210R), and TEL-JAK2 using RetroNectin according to the manufacturer’s instructions (Takara Bio Inc., Shiga, Japan). These cells were cultured in RPMI-1640 medium (Nacalai Tesque, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS) (BioWest, Nuaillé, France), 100 units/ml penicillin (Nacalai Tesque), 100 μg/ml streptomycin (Nacalai Tesque), 2 ng/mL IL-3 (PEPROTECH), and 5 μg/mL puromycin (InVivoGen).

The CML-derived cell line K562 and murine pro-B cell line Ba/F3 were purchased from the Riken Cell Bank (Ibaraki, Japan). Ba/F3 cells expressing p190^BCR-ABL and its point mutant (T315I) were established as previously described [41 (link)]. K562 cells and transduced Ba/F3 cells were cultured in RPMI 1640 (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (Biowest, Nuaillé, France), 100 units/mL penicillin (Nacalai Tesque), and 100 μg/mL streptomycin (Nacalai Tesque). K562 cells were infected with an empty virus and a retrovirus to express BRAF (V600E) using RetroNectin (Takara Bio Inc., Shiga, Japan), and infected K562 cells were selected using culture media supplemented with 2 μg/mL puromycin (InvivoGen, San Diego, CA, USA).