(A) The heatmap shows the average mRNA expression of ZFP91, and the violin plot shows the distribution of ZFP91 mRNA expression in different cell types from 7 scRNA-Seq data sets for COAD. B, B cells; CD4 Tconv, CD4+ conventional T cells; CD8 T, CD8+ T cells; CD8 Tex, exhausted CD8+ T cells; Mast, mast cells; Mono/Macro, monocytes and macrophages; NK, natural killer cells; Mki67+ T, proliferating Mki67+ T cells; pDC, plasmacytoid DCs. (B) Spearman’s correlation of mRNA expression of Zfp91 and the GSVA score for negative regulation of T cell proliferation in TCGA COAD database. (C and D) GSEA of the signature genes for the regulation of CD8+ αβ T cell proliferation (C) and αβ T cell activation (D) in ZFP91-expressing and ZFP91-silenced T cells. NES, normalization enrichment score. (E and F) qRT-PCR analysis of genes associated with T cell proliferation (E) and activation (F) in tumor-infiltrating T cells from CRC. The normalized Zfp91 expression value of tumor-infiltrating T cells with the lowest expression of Zfp91 was set at 1. The normalized Zfp91 expression values of Zfp91hi T cells were higher than 2 (n = 4), and those of Zfp91lo T cells were less than 2 (n = 6). Data in E and F are representative of 3 independent experiments. Data are represented as the mean ± SEM. *P < 0.05 and **P < 0.01 by calculated by permutation test (C and D) and 2-tailed Student’s t test (E and F).