Human chronic myeloid leukemia (CML) in blast crisis cell line (K562) was procured from the American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained at 37°C, 5% CO2 in complete medium, containing Iscove’s Modified Dulbecco’s Medium (IMDM), 1X penicillin/streptomycin (Gibco), and supplemented with 10% (v/v) fetal bovine serum (FBS).
To differentiate K562 into Mk, cells were treated with Phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich, St. Louis, MO, USA), gilteritinib, quizartinib, dubermatinib (TP-0903), and midostaurin for 48–72 h as previously described (15 (link)). Where indicated, low-serum medium supplemented with 1% FBS and 1 μM imatinib were used.
gilteritinib, quizartinib, dubermatinib (TP-0903), and midostaurin were from Cayman chemical (Ann Arbor, MI, USA). All small molecules were dissolved in DMSO and store in −20°C. pMSCV-Flt3-ITD-Y591F/Y919F was procured from Addgene (Addgene.org). Flt3-ITD-Y591F/Y919F cassette was subcloned into lentiviral vector to produce viral particles using 293T cell.
To differentiate K562 into Mk, cells were treated with Phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich, St. Louis, MO, USA), gilteritinib, quizartinib, dubermatinib (TP-0903), and midostaurin for 48–72 h as previously described (15 (link)). Where indicated, low-serum medium supplemented with 1% FBS and 1 μM imatinib were used.
gilteritinib, quizartinib, dubermatinib (TP-0903), and midostaurin were from Cayman chemical (Ann Arbor, MI, USA). All small molecules were dissolved in DMSO and store in −20°C. pMSCV-Flt3-ITD-Y591F/Y919F was procured from Addgene (Addgene.org). Flt3-ITD-Y591F/Y919F cassette was subcloned into lentiviral vector to produce viral particles using 293T cell.