MC38 colon adenocarcinoma cells were cultured in DMEM (Dulbecco’s modified essential medium) (10% FBS, 1% penicillin/streptomycin). Mice were inoculated subcutaneously with 5 × 105 MC38 cells in the right flank. Tumor-infiltrating lymphocytes were prepared by mincing and digesting tumor tissues in collagenase IV (1 mg/ml; Roche) and DNase I (200 U ml−1; Sigma-Aldrich) for 1 hour at 37°C, followed by Percoll density gradient centrifugation.

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