Fig. 3

EV-encapsulated miR-101 from HUCMSCs downregulates BRD4 expression in the HTR-8/SVneo cells. a Intersection of downstream regulatory genes of miR-101 predicted by TargetScan, RNAInter, and StarBase with genes related to eclampsia in the GeneCards database. b Enrichment result of candidate genes by GO analysis from the online KOBAS. The x-axis indicates the number of genes involved, while the y-axis indicates the GO entry name. The dot color indicates −log10 p value, and the dot size indicates the GO entry background gene number. c Binding relationship between miR-101 and BRD4 predicted by bioinformatic analysis. d, e mRNA and protein expression of BRD4 in the placental tissues in PE patients and healthy donors determined by RT-qPCR and immunoblotting. f Correlation between miR-101 and BRD4 in PE determined by Pearson’s correlation analysis. g The targeting relationship between miR-101 and BRD4 determined by dual luciferase reporter gene assay. h The targeting relationship between EV-encapsulated miR-101 and BRD4 determined by dual luciferase reporter gene assay. i The transfection efficiency of miR-101 in HTR-8/SVneo cells and in EVs isolated from HUCMSCs infected with miR-101 mimic. j The transfection efficiency of EV-encapsulated miR-101 in HTR-8/SVneo cells and in EVs isolated from HUCMSCs infected with miR-101 mimic. k, l The mRNA and protein expression of BRD4 after miR-101 or EV-encapsulated miR-101 treatment. *p < 0.05 vs. **p < 0.01. WT, wild-type 3′-UTR BRD4; MUT, mutated type 3′-UTR BRD4; M.NC, negative control mimic; M.miR-101, microRNA-101 mimic; I.NC, negative control inhibitor; I.miR-101, miR-101 inhibitor