Fig. 2: Single-cell analysis reveals the heterogeneity of tumor cells and CAFs in PDAC with Schwann cell accumulation.

a Representative H&E images of PDAC tissues with neural hypertrophy. asterisk, nerves; arrow, stroma; triangle, cancer cells. Scale bar, left, 100 µm; right, 50 µm. b Representatives of IHC staining images of patient A with anti-PGP9.5, GFAP, and S100β. Scale bar, 50 µm. a, b Images are representative of four PDAC samples with similar results. c t-SNE plot depicting the major cell types identified by single-cell RNA sequencing of four PDAC tissues. d UMAP plot showing two main subclusters, basal-like and classical tumor cells from PDAC tissues. e GO analysis showing the upregulated pathways in basal-like and classical tumor cells. f The differentiation states of cancer cells defined by CytoTRACE. For each boxplot, the first quartile, median, and third quartile ± s.d were displayed. n = 4 PDAC patients. g OS analysis of patients with low (n = 44) or with high (n = 44) basal-like signature in the TCGA-PAAD cohort (n = 88). h UMAP plot showing three main subclusters of fibroblasts, including iCAFs, myCAFs, and stellate-like cells. i GO analysis showing the upregulated pathways in iCAFs, myCAFs, and stellate-like cells. j Trajectory of fibroblasts along pseudotime in a two-dimensional space. k Expression of NGFR in the two cell fates of fibroblasts along pseudotime. The expression of NGFR was determined by branch expression analysis. l Heatmap showing the dynamic changes of gene expression along pseudotime. The differentially expressed genes were clustered hierarchically into three groups and the representative enriched pathways of each group were shown. Statistical analysis: one-sided Fisher’s exact test (e, i); two-sided Wilcoxon rank-sum test (f); log-rank test (g).