Fig. 8: Single-cell RNA sequencing analysis shows changes of endogenous Foxf1 mRNA in FEL1^−/− and FEL4^−/− lung cells derived via blastocyst complementation.

a The integrative single cell RNAseq analysis of tdTomato+ chimeric lung cells derived from control mESC s(WT), FEL1^−/− mESCs (MT1), and FEL4^−/− mESCs (MT4) via blastocyst complementation. The cells from the three datasets were grouped into six cell clusters based on mRNA signatures: Endothelial, Epithelial, Fibroblast, Myofibroblast, Matrixfibroblast and Pericyte. b DotPlot shows mRNAs used as representative markers for each cell cluster. c The unsupervised clustering and UMAP projection of single cells from each individual WT (7131 cells), MT1 (4512 cells) and MT4 (6590 cells) datasets are shown. d VlnPlot analysis indicates that Foxf1 mRNA is decreased in MT1 endothelial cells as well as in MT4 pericytes and fibroblasts (upper panel). The box in the box plot represents the middle 50% of the data, with the lower and upper edges of the box indicating the first and third quartiles, respectively. The line inside the box represents the median. The whiskers extend from the box to the minimum and maximum values. The average Foxf1 expression in each cell type is indicated in bottom panels for endothelial cells (WT:2722), (MT1:296) and (MT4:2543); fibroblast (WT:978), (MT1:615) and (MT4:646); pericyte (WT:1363), (MT1:783) and (MT4:1937). The numbers following each genotype indicated the cells used to calculate Foxf1 expression level in each genotype from single cell RNA sequencing data. One-way analysis of variance (ANOVA) analysis was used to determine statistical significance. Multiple means were compared using one-way ANOVA with the post-hoc Tukey’s test. The pvalues for endothelial with significant difference: 0.00535, 0.00856, 0.01851; The pvalues for pericyte with significant difference: 0.00643, 0.00518; The pvalues for fibroblast with significant difference: 0.00406, 0.00313. p < 0.01 is **p < 0.05 is *, ns is not significant.