Crl 1596

Raji (CCL-86; ATCC) and Ramos (CRL-1596; ATCC) were grown as monolayer cultures in RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum (Gibco), penicillin (100 units/mL), streptomycin (100ug/mL), and l-glutamine (200 mM) at 37 °C in a humidified, 10% CO₂ incubator. Raji and Ramos tested negative by PCR for mouse viruses and mycoplasma. Cells were authenticated by STR profiling and were 100% matched to the database.

All the human cell lines [H460 (ATCC^® HTB-177, Manassas, VA, USA); A549 (ATCC^® CCL-185); Caco-2 (ATCC^® HTB-37™); HT-29 (ATCC^® HTB-38™); MCF-7(ATCC^® HTB-22); MDA-MB-231 (ATCC^® HTB-26); Jurkat, T-cell leukemia (DSMZ ACC 282, Braunschweig, Germany) and Ramos B-cell lymphoma (ATCC^® CRL-1596™)] were cultured at 37 °C in a humidified CO₂ incubator (5% CO₂) in complete RPMI media (hematological cell lines Jurkat and Ramos) or DMEM media (adherent cells H460, A549, Caco-2, HT-29, MCF-7, and MDA-MB-231) (both supplemented with 10% (v/v) FBS and 1% (v/v) penicilin–streptomycin). When the cells reached 80% confluence, they were subcultured. For this purpose, adherent cells were washed with PBS and treated with 1 mL 0.25% trypsin-EDTA to detach them. Once collected, cells were centrifugated at 1200 rpm for 5 min. Suspension cells were collected directly from the cell culture plate and centrifugated in the conditions mentioned above. When necessary, the cells were counted using a Neubauer counting chamber and dyed with Trypan Blue.

The human Burkitts Lymphoma cell line Ramos (CRL-1596™) and human T lymphoblast cell line Jurkat (Clone E6-1, TIB-152™) were purchased from ATCC. Low passage (20<) Ramos cells was cultured in complete RPMI 1640 Medium (#A1049101, ATCC modification, ThermoFisher Scientific, Waltham, MA, USA), supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin (15140122, ThermoFisher Scientific), and 10% fetal bovine serum (FBS, A3160801, ThermoFisher Scientific. Low passage of Jurkat cells (20<) was cultured in the same complete medium but without antibiotics. Cells were sub-cultured at 5x10⁵ cells/ml and maintained at 37°C in a humidified 5% CO₂ atmosphere. Recombinant human IL-2 (#11848-HNAH1-E), IL-4 (#11846-HNAE), IL-7 (#11821-HNAE), IL-9 (#11844-H08B), IL-15 (#10360-HNCE) and IL-21 (#10584-HNAE) were obtained from Sino Biological (Beijing, China). JAK inhibitors Tofacitinib (Tof, #4556), Ruxolitinib (Rux, #7064) and Ritlecitinib (Rit, #6506) were purchased from Tocris (Abingdon, United Kingdom). AffiniPure Goat Anti-Human IgM, Fc5μ fragment specific (#109-006-129) was obtained from Jackson ImmunoResearch (West Grove, USA).