Fig. 1

myCAFs enhanced CRC cell stemness and promote migration. A Uniform manifold approximation and projection (UMAP)plot showing the eight major cell types. Dots represent individual cells and colors represent different cell populations. (n = 18 patients). B Single cell data clustering analysis, data from GSE16655 and GSE178318. (n = 18 patients). C Proportion of patients with different Stages in high myCAF and low myCAF score groups, data obtained from the metaGEO database. (n = 913 patients). D Kaplan‒Meier survival curve of patients with high myCAF score and low myCAF score, data obtained from the metaGEO database. (n = 913 patients). E Workflows of the procedure for extracting myCAF and PDOs from CRC Patients and assessing invasion capability after co-culture with CRC Cells. F Western blot analysis of α-SMA, FAP and Collagen I protein expressions in NFs and myCAFs isolated from CRC patients. (myCAF and NF were isolated from four patients). G Immunofluorescence analysis of α-SMA, FAP and Collagen I protein expressions in NFs and myCAFs isolated from CRC patients. Scale bars = 100μm (left), Scale bars = 20μm (right). (n = 3, one of three biological replicates). H Representative α-SMA staining in CRC tissues from FUSCC cohort 2. Scale bars = 50μm. (n = 3, one of three biological replicates). I, J Transwell assay to evaluate the invasion after co-cultured DLD1 with myCAFs. Scale bars = 100μm. (Representative images are shown, data are from five biologically replicates). K, L Sphere forming assay to evaluate the stemness after co-cultured DLD1 with myCAFs. Scale bars = 100μm. (Representative images are shown, data are from five biologically replicates). M, N PDOs 3D invasion assay to evaluate the invasion of PDOs after co-cultured PDOs with myCAFs. Scale bars = 50μm. (Representative images are shown, data are from three replicates). O, P Representative images of subcutaneous tumors treated with FAPI gavage. (n = 5 mice per group). Q, R Liver metastasis assays were performed in vivo by splenic injection to evaluate the effect of FAPI on tumor metastasis. (n = 5 mice per group). Data are shown as mean ± s.e.m. For B-C, data were analyzed by chi-square test. For D, data were analyzed by log-rank test. For I-R, data were analyzed by two-tailed Student’s t-test. P values. *P < 0.05, **P < 0.01, ***P < 0.001