Fig. 7

Pseudotime trajectory analysis assessing the cell fate of fibroblasts in asthma. (A) Pseudotime trajectory analysis to construct fibroblast to ASMC development trajectory. (B) RNA velocity analysis delineated dynamic changes in cell fate, projected onto a UMAP plot. Arrowheads indicated the predicted direction of cell development, with arrow size denoting the strength of predicted directionality. In the NC group (left), cells tend to move from ASMC to Col14a1 + Fib/Myofib, while in the AS group (right), there is a shift from Col14a1 + Fib/Myofib to ASMC. (C) Heatmap showing DEGs along the trajectory-based pseudotime from Col14a1 + Fib/Myofib through Myo/SMC to ASMC. The left-colored bars represented gene cluster C1 to C3. (D) Functional enrichment analysis revealing KEGG pathways and GO terms significantly associated with gene cluster C1 to C3, as depicted in the left figure; the two panels referred to the log10(FDR) and gene count ratio, respectively. (E) Dynamic changes in the expression level of selected DEGs along pseudotime. (F) Violin plots illustrating expression level of selected genes in NC (grey) and AS group (red). (G) Representative immunofluorescence image demonstrating the co-localization of top: POSTN and α-SMA (ACTA2) and bottom: CHRM2 and α-SMA in NC and AS groups