Molm 13

T cell non-Hodgkin’s lymphoma cell line KARPAS299 was supplied by the European Collection of Authenticated Cell Cultures (ECACC). Multiple myeloma cell lines, MM.1S and RPMI8226; Burkitt’s lymphoma cell lines, Daudi, Raji, and NAMALWA; the acute myelogenous leukemia cell line, KG1; the acute T cell leukemia cell line, Jurkat; and the chronic myelogenous leukemia cell line, K562, were supplied by American Type Culture Collection (ATCC). Acute monocytic leukemia cell lines, THP-1, MOLM-13, and NOMO-1; and the myelodysplastic syndrome cell line, SKM-1, were supplied by Japanese Collection of Research Bioresources Cell Bank (JCRB). All these cell lines were maintained in RPMI 1640 (Gibco BRL) supplemented with 10% heat-inactivated fetal bovine serum (Sigma), 100 µg/mL streptomycin, and 100 U/mL penicillin.

The human AML cell lines, MOLM-13, MOLM-14, and KG-1 cell lines were obtained from JCRB Cell Bank. The HL-60 and THP-1 cell lines were purchased from ATCC. The human CML cell line, K562, was obtained from RIKEN BRC Cell Bank. The authentication of purchased cell lines was performed by the respective cell bank, and further authentication was not conducted. The experiments utilized frozen stocks of cell lines, which were prepared within a few passages after acquisition. A Mycoplasma test was not conducted upon receipt from the cell bank. All cell lines were cultured in RPMI1640 medium supplemented with 10% FBS. In the repopulation assay, cells were washed twice with fresh culture medium at the indicated time periods of drug treatment. Then, viable cells were counted using trypan blue staining method and replated in 96-well plates (10⁴ viable cells/well). Cell proliferation was determined using a cell counting kit-8 (DOJINDO) at the indicated timepoints until confluency was reached. In certain experiments, cells cultured in low-adherent culture dishes (EZ-BindShut, IWAKI) to inhibit the adhesion of differentiated leukemia cells were subjected to each analysis.