Fig. 5: Ratio of CD26− and CD26+ fibroblasts change during tumor development and both CD26− and CD26+ NFs contribute to CAF populations. | Nature Communications

Fig. 5: Ratio of CD26− and CD26+ fibroblasts change during tumor development and both CD26− and CD26+ NFs contribute to CAF populations.

From: CD26-negative and CD26-positive tissue-resident fibroblasts contribute to functionally distinct CAF subpopulations in breast cancer

Fig. 5

A UMAP plot of single-cell transcriptomics data of NFs and CAFs isolated during mammary tumorigenesis in WEPtn mice (Fig. 3A) showing CD26 (Dpp4) expression. B Whole mount analysis of normal mammary gland stained for CD26 (in magenta) and smooth muscle actin (SMA, in green) and dapi (in blue). Scale bar is 50 um. Representative image is shown from two biological replicates with similar results. C Flow cytometry analysis of normal mammary gland and WEPtn-derived ILC gated on fibroblasts (EpCAM−/CD45−/CD31− cells) shows the presence of CD26− and CD26+ fibroblasts. Shown in blue is fluorescence-minus-one (FMO) control and shown in red is full stained sample. FMO control was used to set gates. D Percentage of CD26− and CD26+ fibroblasts in control mammary glands (n = 12 mice) and end-stage tumors derived from WEPtn (n = 6 mice), WEH1047R (n = 6 mice), WB1P (n = 7 mice) and WB1P-Myc mice (n = 5 mice). A two-way ANOVA with Bonferonni multiple comparison correction was used to determine statistical significance by comparing CD26− or CD26+ NFs of control to the CD26− or CD26+ CAFs of the different mouse models. Individual data points reflect biological replicates (mice) with mean ± SEM. Source data are provided in source data file. E Partition-based graph abstraction (PAGA) trajectory analysis of single-cell transcriptomics data from Fig. 3A indicates trajectory of NF clusters toward CAF clusters. Pseudotime was plotted using CD26− NFs as a starting point (root) and CD26+ NFs as a starting point. F Clustree analysis of single-cell transcriptomics of fibroblasts during tumorigenesis at various resolutions to show functional relationships between clusters. Box indicates resolution of 0.3 used to analyze the data resulting in 4 clusters: CD26− NFs (cluster 1), iCAFs (cluster 2), CD26+ NFs (cluster 3) and myCAFs (cluster 4). Middle panel and right panel show myCAF and iCAF gene expression signature plotted on clustree, respectively. Light blue indicates high iCAF/myCAF score, dark blue indicates low score.

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