Cell Culture Protocols for HEK293T, Plat-E, and EL4 Cells

HEK293T cells (Cat#CRL-3216, ATCC, VA, USA), were grown in DMEM supplemented with 1% l-glutamine, 1% non-essential amino acids, 1% sodium pyruvate, 1% penicillin/streptomycin, 0.01 M HEPES, 55 μM 2-mercaptoethanol, and 10% FBS (10% DMEM). Platinum-E (Plat-E) retroviral packaging cells (Cat#RV-101, Cell Biolabs, CA, USA) were grown in 10% DMEM. EL4 cells (Cat# TIB-39, ATCC, VA, USA) or male and female primary CD4⁺ T cells, isolated as above, were cultured in RPMI-1640 supplemented with 1% l-glutamine, 1% non-essential amino acids, 1% sodium pyruvate, 1% penicillin/streptomycin, 0.01 M HEPES, 220 μM 2-mercaptoethanol, and 10% FBS (10% RPMI). Generation of MALT1KO Jurkat cells was performed as described^{40 (link)}. All mammalian cell cultures were grown at 37 °C and 5% CO₂. Escherichia coli strain MAX Efficiency DH5α competent cells (Cat#18258012, Invitrogen, Thermo Fisher Scientific, PA, USA) were used for plasmid transformation and preparation. Transformed E. coli DH5α were grown at 37 °C and 230 rpm shaking.