Fig. 5

Transcriptional landscape heterogeneity of LMOD1. A Two-dimensional plots illustrating the invariabilities in LMOD1 expression during the transitions (from normal cells to adenoma cells) along the pseudotime. B Two-dimensional plots showing the variations (decrease) in LMOD1 expression during the transitions (from normal/adenoma cells to cancer cells) along the pseudotime. C Two-dimensional plots indicating the variations (increase) in LMOD1 expression and fibroblasts activation markers during the transitions (from NFs to CAFs) along the pseudotime. D LMOD1 is involved in transdifferentiation from normal epithelium to colorectal cancer but not from normal epithelium to intestine adenomas (grey⊥stands for no change; purple↓stands for decrease). E, F Double immunofluorescence (dIF) staining images of EPCAM (green) and LMOD1 (red) in a resected normal colon specimen (E) and a resected colon cancer specimen (F) (blue, DAPI). LMOD1 is upregulated in normal epithelial cells but downregulate in cancer cells. Scale bars are provided on the graph. G, H Co-localization was determined using the Pearson correlation coefficient in normal colon specimen (G, R = 0.5367, P < 0.0001) and colon cancer specimen (H, R = − 0.5860, P < 0.0001), respectively. The X-axis represents each pixel point on the image, and the Y-axis represents the gray value corresponding to each pixel point. The co-localization relationship between LMOD1 and EPCAM was weaker in tumor tissue compared to that of normal tissue, just like TJs. I dIF staining images of ACTA2 (green) and LMOD1 (red) in a resected normal colon specimen (blue = DAPI), and LMOD1 downregulated in NFs. Scale bars are labelled on the graph. J mIF staining images of ACTA2 (green), LMOD1 (red), and CK (pink) in a resected colon cancer specimen (blue = DAPI), and LMOD1 is upregulated in cancer mesenchymal tissues. Scale bars are provided on the graph. K, L Co-localization was determined using the Pearson correlation coefficient in normal colon specimen (K, R = 0.7930, P < 0.0001) and colon cancer specimen (L, R = 0.9290, P < 0.0001), respectively. The co-localization relationship between LMOD1 and ACTA2 was stronger in tumor tissue compared with that in normal tissue, simile to GJs. The X-axis represents each pixel point on the image, and the Y-axis represents the gray value corresponding to each pixel point. M, N Spatial transcription sections showing the spatial expression of LMOD1 in normal colonic tissue (M) and colon cancer tissue (N). The dot color represents the expression level of the markers. Green boxes for the parenchyma and pink boxes for the mesenchyme. O LMOD1 exhibited similar behaviors as TJs during the malignant transformation of epithelial cells and to GJs in malignant transformation of fibroblasts. P Spearman correlation between LMOD1 expression and the tumor purity (left) as well as infiltration level of fibroblasts in Colon adenocarcinoma (COAD) (middle) and Rectum adenocarcinoma (READ) (right) was analyzed on TIMER 2.0 (TCGA-CRC). Q Spearman association of LMOD1 expression with stromal score (left), immune score (middle), and estimate score (right) was analyzed by “ESTIMATE” package (TCGA-CRC). R, S Spearman association of LMOD1 with fibroblast activation markers ACTA2 (R) and FAP (S) expression (TCGA-CRC). T, U Double staining technique by ACTA2 (green), and FAP (red) staining in the primary CAFs. Representative images of staining are shown. All assays were conducted thrice, independently (scale bars = 20 µm). ANOVA was applied. ****P < 0.0001