Extended Data Fig. 4: Integration of proximal and distal single cell transcriptomes defines the unique gene signature and location of human RASCs.

A) Distal and proximal scRNA-sequencing results from the same patient were concatenated, epithelium was isolated and re-clustered, and cell types were identified based on known markers. B) Distal and Proximal epithelium contribution to the resultant UMAP. C) Expression of SCGB1A1 and SCGB3A2. Secretory cell cluster outlined in yellow, RAS cell clustered outlined in Red. D) Violin plots showing expression of selected genes, highlighting unique genes for each cluster, and shared gene expression between secretory and RAS cell clusters (orange box) and RAS cell and AT2 cell clusters (green box). E) Feature plots of both the proximal and distal epithelium highlighting key gene expression patterns used to identify known cell subtypes. F) Contribution of proximal and distal samples to each cell type, demonstrating that some groups are unique to each region while some are shared. G) Heat map showing gene expression of Secretory, RAS cell, and AT2 cell gene signatures from proximal and distal concatenated data set. Of note, these pairwise comparisons were done between the displayed groups only, as such the resultant gene expression is distinct from the cluster-defining genes we observed in Extended Data Fig. 2a, where the comparisons were done between all epithelial populations. Origin of cells shown in blue (proximal) and orange (distal) bar across top of heat map.