Fig. 3: CAF^{WPOI 4-5} type is characterized by high OXTR expression.

a Differential gene analysis on enriched KEGG pathway from RNA sequencing of CAF^{WPOI 1-3} (n = 3 independent CAFs) and CAF^{WPOI 4-5} (n = 9 independent CAFs). Individual gene expression is shown as in a heatmap. b OXTR protein and mRNA expression in primary CAFs. P = Student’s t test. The experiments were performed three times with similar results. c Correlative analysis between OXTR gene expression and other stromal cell markers in HNSCC, n = 530 (TCGA dataset, Firehose Legacy). d Graphical representation of OXTR mRNA expression in CAFs or tumor cells from breast cancer (CAF: n = 3, tumor cell: n = 4) and PDAC (CAFs: n = 3, tumor cells: n = 2), P = two-tailed t test. e Graphical representation of OXTR mRNA expression from flow cytometry-sorted EpCAM⁺ tumor cells, CD45⁺ leukocytes, CD31⁺ endothelial cells, PDGFR-β⁺ or FAP⁺ stroma fibroblasts. P = two-tailed t test. f Representative fluorescence in situ hybridization in ACTA2-cy3 and OXTR-fam-labeled stroma fibroblasts (scale bar: 20 μm) and (g) IF staining of OSCC tissues with WPOI 1–3 or 4–5 type indicated the localization of OXTR⁺ CAFs (scale bar: 100 μm) and (h) relative quantitation (n = 10 independent samples for each group), P = two-tailed t test. i Representation of GSVA gene set analysis of the oxytocin signaling pathway performed on human HNSCC scRNA-seq datasets (GSE103322). The similarities between different fibroblasts subtypes and myCAFs, iCAFs and apCAFs were analyzed in the lower bubble plots. j Representative IHC analysis of staining for OXTR⁺ CAFs from OSCC patients with WPOI 1–3 (n = 56) or 4–5 (n = 138) and (k) their graphical quantification, n = 194. Scale bar: 20 μm (left), 100 μm (middle), 50 μm (right). l Graph representing the correlation between OXTR⁺ CAFs and OSCC patient postoperative recurrence and survival. P = Pearson’s correlation test and Log-rank (Mantel–Cox). n = 193, scale bars, 50 μm. Results are shown as mean and standard deviation (SD). Boxes indicate the first and third quartiles, bands indicate medians, and whiskers indicate ±1.5 interquartile range. Source data are provided as a Source data file.