Figure 4.

Single-cell transcriptomes of actively differentiating PDCs reveal acquisition of an epithelial-to-mesenchymal transitional phenotype in CF PDE cultures

(A) WT and CF PDE on day 2 (D2), day 5 (D5), day 7 (D7), and day 9 (D9) of differentiation at ALI underwent 10× single-cell RNA sequencing (scRNA-seq). Shown are uniform manifold approximation and projections (UMAPs) of cell types at each time point of differentiation; cell markers for classification used a combined mouse and human scRNA-seq pancreatic datasets.

(B) Heatmaps of DEGs in CF relative to WT PDE cultures at each time point. Genes relevant to epithelial-to-mesenchymal transition (EMT), diabetes, and cystic fibrosis are highlighted. Time point of differentiation is shown in the color-coded panel on top of each heatmap (using the legend shown in C).

(C) Violin plots of DEGs found in WT vs. CF PDE cultures depicting Log2(TPM) expression level at each time point of differentiation.

(D) Putative upstream regulators of the DEGs in CF PDE cultures. Activation Z score of candidate regulators is shown at each time point of differentiation. Z scores >2 indicate activation (dotted lines) and <−2 indicates inhibition.

(E) Activation Z scores of IPA pathways altered in CF relative to WT PDE cultures at each time point of differentiation.