Fig. 4: Investigating the role of TEAD1 in EVT differentiation.

a Experimental scheme to assess the requirement of TEAD1 during EVT and STB differentiation. b Phase contrast images of H9 WT EVTs and TEAD1 KO hTSCs that have undergone EVT differentiation. The scale bars indicate 75 μm. The images are representative of four independent experiments. c The relative number of invading cells following Matrigel invasion assay of H9 WT EVTs and TEAD1 KO hTSCs that have undergone EVT differentiation. Error bars indicate ±1 standard error of five technical replicates. The center of the error bar indicates the mean. One-tailed student’s t test was used for statistical analysis.‘*‘ indicates a p-value <0.05. The exact p-values were 0.028856521 (#1), 0.020949253 (#2), and 0.029918938 (#3). d Principal component analysis (PCA) of WT and TEAD1 KO hTSCs, EVTs, and STBs based on RNA-seq data. e Scatter plot showing the differential gene expression analysis between H9 WT and TEAD1 KO EVTs. WT contains two RNA-seq samples, and TEAD1 KO contains two RNA-seq samples each from three independent clones. f Selected GO biological processes that are enriched among DEGs significantly upregulated or downregulated in TEAD1 KO “EVTs” relative to WT EVTs. g The expression of EVT-specific genes (n = 462)³⁷ in H9 WT and TEAD1 KO EVTs. WT contains two RNA-seq samples, and TEAD1 KO contains two RNA-seq samples each from three independent clones. Two-tailed Wilcoxon Rank Sum Test was used for statistical analysis. Boxplot presents the 25th, median, and 75th quartiles, the whiskers extend 1.5 of interquartile ranges. h Principal component analysis (PCA) of WT and TEAD1 KO hTSCs, EVTs, and STBs based on ATAC-seq data. i Selected GO biological processes that are enriched among DARs significantly more open or closed (blue) in TEAD1 KO “EVTs” relative to WT EVTs. j WT and TEAD1 KO EVT RNA-seq and ATAC-seq data shown in the vicinity of selected genes. k Levels of WT hTSC and WT and TEAD1 KO EVT ATAC-seq signal over regions that are specifically open in WT EVTs. Each condition contains two ATAC-seq samples.