Fig. 5. Collagen XII is produced by cancer-associated fibroblasts in mouse and human breast tumours.

a UMAP visualisation and clustering of 11,490 cells analysed by single-cell RNA-sequencing from Valdes-Mora et al.⁶⁶ of late-stage mammary tumours. b Feature plot and c Violin plot showing the log-normalised expression of Col12a1 over the UMAP structure and clusters as shown in a confirming the source of collagen XII expression. d UMAP visualisation and clustering of 100,064 cells analysed by single-cell RNA-Sequencing from the Breast Cancer Atlas⁶⁷, a study of 26 human primary breast cancers. Cell clusters are labelled by their major lineage annotations for B cells, Plasmablasts, T cells, Myeloid, Cancer epithelial, Normal epithelial, cancer-associated fibroblasts (CAFs), perivascular-like cells (PVL) and endothelial cells. e Feature plot and f Violin plot showing the log-normalised expression of COL12A1 over the UMAP structure and major lineage clusters as shown in d. g Western blot analysis of collagen XII secretion by normal mammary fibroblast (NF) and cancer-associated fibroblasts (CAFs) (*denotes non-specific band). GAPDH is loading control. Representative of n = 3 biologically independent experiments. h Quantification of Col12a1 mRNA expression (n = 3 biologically independent experiments ****p < 0.0001 one-way ANOVA with Tukey’s multiple comparison test) and i collagen XII protein expression (representative of n = 3 biologically independent experiments) in collagen XII knockdown CAF lines generated using collagen XII short-hairpin mRNA scrambled control (shScr) and four collagen XII targeting constructs (shColXII#1–#4) (*denotes non-specific band). Vinculin is loading control. Source data are provided in the Source data file.