Fig. 5

Hsa_circ_0002348 regulated trophoblast proliferation and apoptosis by competitively binding to miR-126-3p. A Schematic illustration showing hsa_circ_0002348 binding to five miRNAs predicted by online tools; B CCK-8 assay displaying the augmented trophoblast proliferation after transfected with miR-126-3p mimics; C RIP assay detected by qRT-PCR showing the enrichment of hsa_circ_0002348 pulled down by AGO2 in miR-126-3p mimics treated with HTR8 cells; D colony-formation assays showing the proliferation ability of HTR8 cells treated with miR-126-3p mimics; E flow cytometry of fluorescence analysis stained with Annexin V-FITC and PI detecting the apoptosis rate of HTR8 cells treated with miR-126-3p mimics; F CCK-8 assay showing cell proliferation of HTR8 cells with hsa_circ_0002348 overexpression and miR-126-3p mimics; G flow cytometry of fluorescence analysis stained with Annexin V-FITC and PI detecting the apoptosis rate of HTR8 cells treated with hsa_circ_0002348 overexpression and miR-126-3p mimics; the quantitative histograms of D, E, and G displayed in the right; H western blotting analysis showing the expression level of p-mTOR, mTOR, p-ERK1/2, ERK1/2 when compared with Tubulin as control in HTR8 cells with NC or miR-126-3p mimics; the quantitative analysis of band intensity performed with Image J (right); I representative FISH images of miR-126-3p expression in the pregnant mice’s placentas of each group (n = 3/group); J representative FISH images of miR-126-3p expression in the preeclampsia patients’ placentas (n = 3)and normal healthy controls (n = 3); miR-126-3p signals in green, and DAPI-stained nuclei in blue; Scar bars, 50 μm. circ_0002348 over representing hsa_circ_0002348 overexpression; NC negative control, Ctrl control; the data presented as mean ± SEM; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001