Fig. 5: Synthetic miR-214-3p administration induces PE-like phenotypes in pregnant mice.

Pregnant mice were infused with locked nucleic acid-based control miRNA (C), synthetic miR-214-3p mimics (M), or miR-214-3p plus rmPlGF-2 (PlGF-2) using an osmotic pump from GD 9.5 to GD 17. a Circulating PlGF levels were determined using an ELISA kit (n = 5). b Levels of eNOS were determined in maternal aortic vessels using western blotting and quantified using ImageJ (n = 5). c, d The levels of NO and cGMP synthesis were determined in aortic vessels and their lysates using DAF-FM-based fluorescence microscopy and an ELISA kit, respectively (n = 5). e Vasorelaxation responses of aortic vessels to Ach were determined using myography (n = 5). f The mean blood pressure (BP) was determined using a noninvasive tail-cuff method (n = 5). The arrow indicates surgical implantation of the osmotic pump. g The urine albumin-to-creatinine ratio (UACR) was calculated after determining the levels of these molecules using ELISA kits (n = 5). h Representative images of placental tissues stained with DAPI (nuclear marker) and antibodies against cytokeratin (CK, a trophoblast marker), CD31 (an endothelial cell marker), or α‐smooth muscle actin (α-SMA, a smooth muscle cell marker) and visualized using confocal microscopy. Scale bar = 100 μm. i Average fetal weight at GD 17 (5–10 fetuses/litter, n = 5 litters). The data are presented as the mean ± SEM. Statistical significance was determined using multiple t tests (e, f) or one-way ANOVA (a–d, g, i), followed by the Holm–Sidak’s multiple comparisons test. ns not significant.