Fig. 9

WCGNA-based Transcriptomic Profiling of EVs for Sex and Inter-Pregnancy Differences. WGCNA of RNA transcripts expressed in 16 EV samples, shows that EVs have distinct gene networks and correlations with the trait of ‘Sex’ (female vs. male) contributing to the majority of the difference between clusters. The second trait, ‘Pregnancy’, was a smaller contributor to the overall composition of identified networks. A Topological overlap matrix (TOM) visualizing WCGNA in EVs. The gray module represents residual genes that could not be associated with networks. B Multidimensional Scaling (MDS) plot of genes in EVs show that the ‘turquoise’ and ‘brown’ module eigengenes are most separated from each other. C Module eigengene (ME) dendrogram as the dissimilarity measurement between MEs (1st principal component), showing the highest branching out of ME ‘brown’ away from ME ‘turquoise’, also suggesting the greatest dissimilarity. D Heatmap of WGCNA module correlations with sample traits of ‘Pregnancy’ and ‘Sex’. The results of this analysis show that ‘Sex’ (female vs. male) is the most important determinant of module identity, with the ‘turquoise’ module identifying male, and the ‘brown’, ‘salmon’, ‘black’, and ‘pink’ modules identifying female samples. The ‘turquoise’ and ‘black’ modules were also significantly associated with the trait of ‘Pregnancy’ and represent pregnancy-to-pregnancy variation in gene networks in EVs. E Bar plot of ‘Sex’ (female vs. male) trait-based module significance (average gene significance in a module) across modules in EVs