Human Jurkat T leukemia cells [American Type Culture Collection (ATCC), TIB-152] were cultured in RPMI 1640 (Invitrogen) containing 10% fetal calf serum (FCS; Invitrogen) plus penicillin (10 U/ml) and streptomycin (10 mg/ml) (Invitrogen). HEK293T cells (ATCC, CRL-11268) were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen) containing 10% FCS plus penicillin (10 U/ml) and streptomycin (10 mg/ml). All cell lines used were tested and confirmed to be negative for mycoplasma. Primary murine T cells were negatively selected from the spleen, lymph nodes, or peripheral bloods of mice using magnetically coupled antibodies against CD11b, B220, CD49b, CD235, and TER-119 [magnetic cell separation kit (Miltenyi Biotec)]. To induce IL-17A production of 3-day–cultured T cells (Fig. 1E), the primary T cells were stimulated with biotin-conjugated anti-CD3 antibodies (3 μg/ml) plus streptavidin (3 μg/ml) for 3 hours at 37°C.

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