Fig 1: 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.
Fig 2: Collagen XII expression is associated with poor prognosis in human breast cancer.a COL12A1 expression in human non-tumour mammary tissue (n = 98) or primary tumour (n = 1080) from ‘The Cancer Genome Atlas’ (TCGA) breast adenocarcinoma cohort (BRCA)60 (Median and interquartile range, p value determined by two-sided Mann-Whitney test (****p < 0.0001 Mann–Whitney U-test). b Kaplan–Meier overall survival and c progression-free survival analysis of collagen XII stratified (tertiles) breast cancer patients from TCGA BRCA dataset (n = 1067). Univariate Cox Proportional Hazards Model hazard ratio (HR), HR 95% confidence interval and p value for collagen XII high vs low are indicated. d Representative collagen XII IHC-stained TMA sections from triple-negative human breast cancer (TNBC) cohort61 of n = 150 patients (scale bar = 100 µm). e Kaplan–Meier analysis of disease-specific survival and f distant recurrence from TNBC TMA sections, median-stratified by % stromal collagen XII staining (n = 150, 3 TMA cores per patient). Univariate Cox Proportional Hazards Model hazard ratio (HR), HR 95% confidence interval and p value for collagen XII high vs low are indicated. Source data are provided in the Source data file.
Supplier Page from OriGene Technologies for Col12a1 Mouse shRNA Plasmid (Locus ID 12816)