Fig. 4

Monocle trajectory analysis of epithelial cell subgroups and gene expression dynamics. A–D Monocle trajectory analysis of epithelial cell subgroups, with cells colored by (A) inferred pseudotime, (B) developmental status, (C) group designation, and (D) sample origin. (E) The proportion of each epithelial cell subtype in each state. F Pseudotemporal heatmap displaying expression changes in the top 30 genes identified by BEAM analysis before and after node 1. G Pseudotemporal heatmap revealing the top 30 genes with significant expression shifts along the trajectory. H Gene expression patterns of DCBLD2, GLUL, JUN, and TAGLN2, with color coding based on expression levels throughout the pseudotime progression. I Expression levels of DCBLD2, GLUL, JUN, and TAGLN2 before and after node 1, displayed along pseudotime and color coded by group.(J) Violin plots illustrating the expression of GLUL, JUN, and TAGLN2 in in situ versus metastatic cancers