Fig. 7: Meis1 expression in EC triggers transcriptional changes akin to pre-HE cells.

A Schematic of the workflow used to investigate the effect of Meis1 overexpression in EC (created with BioRender.com). B Mean fluorescence intensity (MFI) of CD44 immunostaining 72 h post-transduction. Transduced EC were selected (VEC⁺CD31⁺YFP⁺) for the experiments (n = 2 parental untransduced CTRL, 3 YFP-CTRL, and 3 Meis1-OE). Bar plot on the right shows the mean ± SEM (p-values based on two-sided t-test). C Volcano plot showing DEG in Meis1-OE EC compared to YFP-CTRL EC by RNA-sequencing (p-values based on Wald test with Benjamini–Hochberg correction). D Heatmap depicting all genes up-/down-regulated in Meis1-OE EC in common with DEG in the CITE-seq analysis of aEC vs pre-HE cells. Hypergeometric test (one-tailed) was performed to determine whether the overlap was greater than expected from random sampling. E GSEA analysis quantifying enrichment of custom pre-HE genesets (based on CITE-seq expression) in Meis1-OE EC (p-values adjusted with Benjamini-Hochberg correction). F–H Transduced cells with Meis1-OE or YFP-CTRL vector were co-cultured on OP9 cells for 6 days followed by immunophenotyping. F gMFI values for the expression of surface markers relevant to EHT subpopulations and (G) percentage of cells expressing each marker. Data are presented as mean ± SEM and each data point depicts the average of technical duplicates (n = 3 independent biological samples for each group). P-values were calculated using a two-sided t-test. H Histogram plot of CD44 expression by flow cytometry after OP9 co-culture. Source data are provided as a Source Data file.