library_construction_protocol: Tumor xenografts were mechanically and enzymatically (10 U/mL collagenase I, 400 U/mL collagenase IV, and 100 μg/mL DNase I in FBS-free RPMI-1640 medium) digested to obtain a single-cell suspension. Intratumoral CD45+T cells were isolated using BD FACSAriaIII and resuspended in PBS containing 0.04% BSA. Cells were counted and cell density was adjusted to that recommended for 10x Genomics Chromium single-cell 3' v3 processing and library preparation. Sequencing was performed on an Illumina platform (NovaSeq 6000), by GENERGY BIO (Shanghai, China), at a sequencing depth of about 90,000 reads per single cell. Sequencing data in a bcl file were converted to FASTQ format using Illumina bcl2fastq2 Conversion Software v2.20 and were subjected to quality control by FastQCv0.11.9.