Research Paper Volume 14, Issue 7 pp 3049—3069

Figure 4. miR323-5p is the downstream target of circGSE1. (A) The downstream miRNA targets of circGSE1 as predicted by the TargetScan, miRanda and RNAhybridB databases. (B) The relative OEC expression of the downstream miRNA target of circGSE1 predicted by the database as determined by qRT-PCR. (C) The relative YEC expression of the downstream miRNA target of circGSE1 predicted by the database as determined by qRT-PCR. (D)The relative of miR323-5p expression in YEC and OEC. (E, F) The FISH experiment showed that most circGSE1 (red) and miR-323-5p (green) are colocalized in the cytoplasm (magnification, 400×, scale bar, 20 μm) (n=3, analysed 8 cells). (G) Schematic diagrams of the circGSE1-WT and circGSE1-mutant dual-luciferase reporter vectors. (H) Dual-luciferase reporter assays showed that the circGSE1 wt 3’UTR but not the mut 3’UTR was downregulated by miR-323-5p mimics or upregulated by the miR-323-5p inhibitor. (I, J) The Anti-Ago2 RIP assay was executed in OEC, followed by nucleic acid electrophoresis and qRT-PCR to detect circGSE1. (The data are expressed as the mean ± SD, *P < 0.05, **P < 0.01 versus the negative control).