Tf 1a

Human myeloid leukemia cells lines, TF-1a, TF-1, and MV4-11, were purchased from ATCC (Rockville, MD) and maintained in RPMI 1640 (TF-1a and TF-1) or IMDM (MV4-11) supplemented with 10% FBS without (TF-1a) or with GM-CSF (5 ng/mL, TF-1, MV4-11) at 37°C in humidified air containing 5% CO₂. All the media and sera were purchased from Life Technologies, Inc. (Gaithersburg, MD). During log-phase growth, the cells were collected and transferred to 6-well plates at a concentration of 10⁵ cells/mL. For differentiation study, PMA (Sigma Chemical Co., St. Louis, MO) was added to the cells and incubated for 48–72 h. For apoptosis assay, cells were treated with Bay 11-7085 (Calbiochem, La Jolla, CA) at different concentrations. The control cells were treated with PMA or Bay 11-7085 solvent. After the treatment for 24 and 48 hours, the cells were harvested into a 15 mL polypropylene centrifuge tube and spun down for 8 min at 600 RPM (80 g), after which the supernatant was discarded and the cells were resuspended in 0.5 mL of culture medium and 1-2 drops of the cell suspension were placed on a slide in the central area and moved around to form a thin and even film with a glass spreader. The glass spreader was made from glass transfer pipette over an alcohol burner for few seconds to minutes.

HEK293T/17 cells (ATCC no. CRL-11268) were cultured in DMEM GlutaMAX medium (GIBCO), K562 (ATCC no. CCL-243), HEL (ATCC no. TIB-180), TF-1a (ATCC no. CRL-2451), Jurkat (ATCC no. TIB-152), and U937 (ATCC CRL-1593.2) cells in RPMI 1640 GlutaMAX medium (GIBCO). Cells were tested as mycoplasma free. The cells were supplemented with 10% fetal calf serum (FCS) and 1% Penicillin/Streptomycin (GIBCO) and cultured at 37 °C in a 5% CO₂ atmosphere.

Human lymphocyte cell line DG75 (ATCC CRL-2625), derived from an Epstein-Barr virus (EBV)-negative primary abdominal B cell lymphoma [40 (link)], was obtained from Paul D. Ling (Baylor College of Medicine). Cell lines BJAB (DSMZ ACC-757), an EBV-negative B-lymphoblastoid cell line [41 (link)], and CEM (ATCC CCL-119), developed from an acute T cell leukemia [42 (link)], were obtained from Linda R. Gooding (Emory University, Atlanta, GA). Human B lymphocyte cell line RL (ATCC CRL-2261), derived from a non-Hodgkin’s lymphoma ascites [43 (link)], and TF-1a (ATCC CRL-2451), a myeloid leukemia cell line that can undergo macrophage-like differentiation [44 (link)], were obtained from the ATCC. Lymphocyte and leukemia cell lines were grown in RPMI medium (Cellgro, Manassas, VA) supplemented with 10% fetal bovine serum (FBS; Hyclone, Logan, UT) and 2 mM glutamine. The TC7 cell line, an African green monkey kidney cell line derived from CV-1 cells that is permissive for SV40 replication [45 (link),46 (link)], was cultured in Gibco Eagle’s minimum essential medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 5% FBS, 10% tryptose phosphate broth, 2% Gibco MEM vitamin solution, and 0.25% glucose [47 (link)].