Hl 60

Cell lines S1T (HTLV-1-infected CD4⁺ T-cell line derived from an ATL patient; kindly provided by Dr. Naomichi Arima, Kagoshima University), [23 (link)] MT-2 (HTLV-1-infected T-cell line derived from normal human leukocytes transformed by leukemic T-cells from an ATL patient) purchased from Japanese Cancer Research Resources Bank (Osaka, Japan; catalogue number: JCRB1210), [24 (link)] Jurkat (T-lineage acute lymphoblastic leukemia cell line) purchased from RIKEN BioResource center (BRC) (Ibaraki, Japan; catalogue number: RBRC-RCB3053), and HL60 (acute myeloid leukemia cell line) purchased from RIKEN BRC (catalogue number: RBRC-RCB0041) were cultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal calf serum, 2 mM l-glutamine, 0.1 mg/mL streptomycin, and 100 U/mL penicillin [8 (link)].

THP-1, HL-60, HEL 92.1.7, U-937, and K562 cell lines were obtained from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). Peripheral blood mononuclear cells (PBMCs) were isolated and prepared from healthy volunteer donors using Ficoll-Paque Plus Reagent (GE Healthcare, Buckinghamshire, UK). These cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc., Rockford, IL, USA) and 1% NEAA in a 5% CO₂ incubator at 37 °C.

Human myeloid leukemia HL-60, mouse macrophage-like J774.1, and HEK293 FT cells were obtained from the RIKEN BioResource Center (Tsukuba, Japan), the American Type Culture Collection (Manassas, VA), and Thermo Fisher Scientific (Waltham, MA), respectively. The generation of HL-60-derived USP2KD cells and their control cells have been described previously [29 (link)]. The USP2KD-derived cells, in which USP2A (USP2AR), USP2B (USP2BR), and isopeptidase-mutant USP2A (C276AR) were reintroduced, have also been previously discussed [29 (link)]. HL-60 cells and their derivatives were differentiated into macrophage-like cells by treatment with 30 nM of phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) for 1 d. Peritoneal macrophages were collected from mice treated with thioglycollate medium (2 mL/head; Sigma-Aldrich). Cells were collected from mouse peritoneal cavities using a 10 mL syringe with an 18-gauge needle; after which, macrophages were separated using an adherence-based method [32 ].

Methylthiazolyldiphenyl-tetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), and propidium iodine (PI) were purchased from Sigma-Aldrich (St. Louis, MO, USA). JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′ Tetra- ethylbenzimidazolylcarbocyanine iodide) was obtained from Molecular Probes (Invitrogen, Karlsruhe, Germany). Q-VD-OPh was purchased from R&D (Minneapolis, MN, USA). The antibodies of GAPDH, actin, PARP-1, and Rad51were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Antibodies of DFF45/DFF35, TNFR1, TNFR2, Fas, DR5, Bid, cleaved caspase-1, caspase-3, caspase-5, caspase-8, gasdermin D, cIAP1, cIAP2, survivin, HMGB1, and γH2A.X^Ser139 were obtained from Cell Signaling Technologies (Boston, MA, USA). Antimouse and antirabbit IgGs were obtained from Jackson Immuno-Research Laboratories, Inc. (West Grove, PA, USA). HL-60 (promyelocytic leukemia) was obtained from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). HL-60 cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) (Gibco, Grand Island, NY, USA) with 10% FBS (v/v) and penicillin (100 U/mL)/streptomycin (100 mg/mL). The cells were grown in a water-saturated atmosphere at 5% CO₂ and at 37 °C.

A human promyelocytic leukemia cell line HL60 was purchased from Riken BRC Cell Bank (Tsukuba, Japan) and cultured in RPMI 1640 (Wako, Mie, Japan) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (Nichirei Biosciences, Tokyo, Japan). The cell concentration of HL60 cells and granulocytes was adjusted to 0.5 × 10⁶ and 4 × 10⁶ /mL in the medium, respectively, for incubation with or without 1 μM lipopolysaccharides (LPS) (Sigma-Aldrich, St. Louis, MO, USA).

Human acute myeloid leukemia (AML) MV4-11 (RRID: CVCL_0064) OCI-AML-2 (RRID: CVCL_1619) cell was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA); the human acute myeloid leukemia THP-1 (RRID: CVCL_0006) and HL-60 (RRID: CVCL_0002) cells were purchased from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan), and the human embryonic kidney HEK293FT (RRID: CVCL_6911) cell was purchased from Thermo Fisher Scientific (Waltham, MA, USA). MV4-11, THP-1, HL-60, and ODC-AML-2 cells were grown in RPMI-1640 Medium (HyClone™, Cytiva, Marlborough, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin. HEK293FT cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) (HyClone™, Cytiva, Marlborough, MA, USA) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. These cell lines were cultured at 37 °C in a humidified incubator containing 5% CO₂. All experiments with cell lines were performed in mycoplasma-free cells.

The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240). The cells were cultured in RPMI-1640 and Iscove’s modified Dulbecco’s medium containing 10–20% fetal bovine serum supplemented with 100 U/mL penicillin/streptomycin (P/S) at 37 °C in a humidified atmosphere of 5% CO₂.