Fig. 2

Gestational variation in EV-associated miRNAs across gestation. EVs were isolated from plasma obtained from women with NGT and GDM across gestation in a cross-sectional study design. Analysis was performed on EVs microRNA profiles generated using next-generation sequencing. A Number of miRNAs that changed significantly across gestational age, and condition (e.g. NGT or GDM). Heatmap of the top statistically significant miRNAs (adjusted P value of < 0.05) across gestation (B), and between NGT and GDM (C), where red and green are high and low expression, respectively. D Linear mixed modelling of 101 statistically significant miRNAs that change across gestation when comparing normal to GDM pregnancies. miRNA counts were normalized using the DESeq2 package in R, before statistical analysis using the likelihood ratio test. Subsequently, linear mixed modeling was performed on the 101 statistically significant miRNAs (p-value < 0.05) that change across gestation when comparing normal to GDM pregnancies, using the lme4 package in R. The data was scaled between 0 and 1, before hierarchical clustering analysis using Euclidean distance, which is displayed as a circular cladogram (generated using the ggtree package in R). E Each color of the circular cladogram represents a different cluster and its trend, as shown in A–T. Within the panels, the color blue is for normal pregnancies whilst the color red is for GDM pregnancies