Figure 5.

The ductal CD133^highCD71^lowFSC^mid-high fraction expresses genes with a progenitor cell signature, co-expressing Sox9, Pdx1, and Nkx6-1. (a) Schematic for droplet RNA-seq (Drop-Seq) analysis. Datasets from CD133^highCD71^lowFSC^low (FSC^low; n = 598) and CD133^highCD71^lowFSC^mid-high (FSC^mid-high; n = 1125) fractions were combined and clustered using Seurat. (b) Average scaled expression (measured by average Pearson residual) of canonical markers for epithelial, pancreatic, and immune cell types. (c) Visualization of clusters using uniform manifold approximation and projection (UMAP) on the combined datasets. (d) The 2 datasets were split in UMAP. (e) Proportion of the FSC^low or FSC^mid-high units found in each cluster from total population. (f–g) Top pathways in cluster 0 or cluster 1, ranked by P-values, identified by gene set enrichment analysis (GSEA) using KEGG database. (h) Units that co-express Sox9, Pdx1, and Nkx6-1 were identified as triple-positive (TP) units and labeled in the UMAP as darker dots (left). Percentage of TP units per cluster is presented (right). (i) IF staining of ductal small clusters. Full arrows indicate Sox9⁺ ductal cells that co-express Pdx1 or Nkx6-1. Arrowheads indicate Sox9^-Pdx1⁺ cells (upper panel) or Sox9^-Nkx6-1⁺ cells (lower panel). Scale bar = 20 µm. (j) Aggregated dataset was first processed using StemID algorithm and the resulting StemID cluster 03 was then back-fitted into the Seurat UMAP clusters 0 to 7 (left). Percentage of StemID cluster 03 units was highest in UMAP cluster 1 (right). Abbreviations: KEGG, Kyoto Encyclopedia of Genes and Genomes.