Jurkat cells

TMNK-1 cells (an immortalized human liver endothelial cell line, JCRB1564), MEG-01 s cells (a human megakaryoblastic leukemia cell line, IFO50473), and THP-1 cells (a human acute monocytic leukemia cell line, JCRB0112) were obtained from the JCRB cell bank (Osaka, Japan). Jurkat cells (a human T lymphocyte cell line, RCB0806) were obtained from the RIKEN BRC. SK-BR-3 cells (an HER2⁺ human breast cancer cell line, ATCC® HTB-30) was obtained from ATCC. Jurkat cells expressing human FcγRIIa or FcγRIIIa with the Nuclear Factor of Activated T cells (NFAT)-driven luciferase reporter (Jurkat/FcγRIIa/NFAT-Luc, Jurkat/ FcγRIIIa/NFAT-Luc) were established previously (14 (link), 15 (link)). TMNK-1 cells were maintained in Medium 200 (Thermo Fisher Scientific, Waltham, MA) supplemented with Low Serum Growth Supplement (Thermo Fisher Scientific) at 37°C in a humidified atmosphere containing 5% CO₂. Other cell lines were maintained in RPMI1640 medium supplemented with 10% FBS at 37°C in a humidified atmosphere containing 5% CO₂. The expression of the FcγRs in the cell lines was measured by FITC-labelled anti-CD64 antibody, anti-CD32 antibody, and anti-CD16 antibody (BD Biosciences; San Jose, CA) using BD FACSCanto II (BD Biosciences).

All experiments were performed using Jurkat cells (an immortalized line of human acute T-cell lymphocyte cells, RIKEN BioResource Research Center, Tsukuba, Japan). Jurkat cells were maintained in RPMI-1640 with l-glutamine and phenol red (FUJIFILM Wako Pure Chemical, Osaka, Japan), 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA), 100 units/mL penicillin, and 100 μg/mL streptomycin (PS, FUJIFILM Wako Pure Chemical) at 37°C, and 5% CO₂. Cells were passaged every 2–3 days and a day before the experiment.

The experiments were performed using Jurkat cells (RIKEN BioResource Research Center, Tsukuba, Japan). Jurkat cells were maintained in RPMI-1640 with l-glutamine and phenol red (FUJIFILM Wako Pure Chemical, Osaka, Japan), 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA), 100 units/mL penicillin, and 100 μg/mL streptomycin (PS, FUJIFILM Wako Pure Chemical) at 37 °C, 5% CO₂. Cells were passed every 2–3 days and a day before the experiment.