Fig. 4

Analysis and verification of ligand-receptor subtype 2 in CRC cell lines and scRNA-seq data. A scRNA-seq data of 29 CRC single-cell samples were clustered according to the subtype marker ligand-receptor pairs. B t-SNE plot of the scRNA-seq data of SMC14, SMC20 and KUL01. C Pathway enrichment analysis of DEGs between subtype 2 and other subtypes of malignant epithelial cells. D GSEA analysis of differential pathways of malignant epithelial cells in scRNA-seq sample. E 3D visualization of the spatial structure of SMC14, SMC20 and KUL01 (right panel). Cell types and density scatter diagrams of longitudinal sections of tumor tissue spatial structure (left panel). F Flow cytometry analysis results of activated CD8 + T-cells. G–J left panel: GSEA analysis of PI3K/AKT, TNFA, TGF-Beta and MAPK pathways in 4 CRC cell lines. G–J right panel: LDH level measured the cytotoxicity of CD8 + T-cell to co-cultured tumor cells in vitro. E:T, effector:target ratio. *P < 0.05, **P < 0.01, ***P < 0.001