Panel A shows a volcano plot of differentially expressed genes in O T I cells. There are 1,297 downregulated genes and 1,085 upregulated genes, with lines indicating a 2-fold difference (D E Seq 2, F D R less than 0.05). Panel B shows G O enrichment dot plot of pathways from differential genes. Panel C shows G S E A plots of T cell signature gene enrichment. Panel D shows heatmaps of stem-like, exhausted, and effector gene expression. Panel E shows flow cytometry plots and quantification of marker expression. Panel F shows flow cytometry plots and quantification across T cell subsets.
Mettl8 deficiency alters transcriptional profiles of CD8 + T cells. RNA-seq analysis were performed using WT or Mettl8−/− (KO) OT-I cells from the adoptive transferred tumor model. (A) Volcano plot illustrates genes differentially expressed between OT-I cells from WT and KO mice. Lines indicate a twofold difference between OT-I cells from KO and WT mice (DESeq2, FDR < 0.05). (B) Gene Ontology (GO) enrichment (biological processes) analysis of DEGs obtained from the R package named clusterProfiler. (C) GSEA shows enrichment of Tcf1+ TPEX signature genes (top), GzmK+ TEM signature genes (middle), and TEMRA signature genes (bottom) in the transcriptome of KO versus WT cells. (D) Heatmaps of DEGs related to stem-like, exhausted, or effector-like features (Log2FC > 1, FDR < 0.05). (E) Representative flow cytometry plots (left) and cumulative data (right) show the expression of the indicated molecules gated on tumor-infiltrating OT-I cells. n = 5–6 per group. (F) Representative flow cytometry plots (left) and cumulative data (right) show the expression of the indicated molecules gated on tumor-infiltrating Tcf1+ Tim3− TPEX, CX3CR1+ Tcf1− Int-TEX, and CX3CR1− Tcf1− TEX subsets. n = 5–6 per group. Data are representative of two independent experiments. P values were calculated by two-tailed Student’s t test; **P < 0.01; ***P < 0.001; ****P < 0.0001.