Fig. 3

The heterogeneous landscape of CC molecules across different lesions in fibroblasts. A Circle plot showing the possible ligand-receptor pairs between fibroblasts and other type cells (predicted by CellChat). B The UMAP plot of all fibroblasts, color-coded for two seurat clusters. C, D The fraction of three tissue types in two cell types showed by histogram (C) and UMAP plot (D). E All fibroblasts were defined as Normal fibroblasts (NFs) and Cancer-associated fibroblasts (CAFs), respectively, according to tissue origin. F Heatmap showing differentially expressed genes between the two cell types (fold change > 1.5, FDR < 0.01). G, H The bubble plots indicate the up-regulated gene set in NFs (G) and CAFs (H), differently. I Trajectory of fibroblasts constructed by Monocle 3. Each point corresponds to a single cell and is colour-coded by pseudotime. J Two-dimensional plots showing the dynamic expression of representative CC molecules during the fibroblast transitions along the pseudotime. K mIF staining images of ACTA2 (pink), CLDN4 (green), and GJA4 (red) in a resected normal colon specimen (blue, DAPI), and GJA4 (for GJs) is lowly expressed in normal mesenchymal tissues (marked by ACTA2). Scale bars are labeled on the graph. L mIF staining images of CK (orange), ACTA2 (pink), CLDN4 (green), and GJA4 (red) in a resected colon cancer specimen (blue, DAPI), and GJA4 is highly expressed in cancer mesenchymal tissues (marked by ACTA2 and CK). Scale bars are labeled on the graph. M, N Co-localization was evaluates based on the Pearson correlation coefficient in normal colon specimens (M, R = 0.2692, P < 0.0001) and colon cancer specimen (N, R = 0.8806, P < 0.0001), respectively. The co-localization relationship between GJA4 and ACTA2 was stronger in tumor tissue compared to normal tissues. The X-axis represents each pixel point on the image, and the Y-axis represents the gray value corresponding to each pixel point. O, P The ultrastructure of junctions in normal colon specimens and colon cancer specimen was examined using a transmission electron microscope (EM). Orange arrows represent TJs, and green arrows represent GJs. Q, R Spatial transcription sections indicate the spatial expression of EPCAM, ACTA2, TJs markers (CLDN3/4/7), and GJs markers (GJA4, GJB1, and GJC1) in normal colonic tissue (Q) and colon cancer tissue (R). The dot color indicates the expression level of the markers. Green boxes for the parenchyma and pink boxes for the mesenchyme