Fig. 5: OGRU interacts with miR-320-3p.

a RNA fluorescence in situ hybridization showed that OGRU primarily localizes in the cytoplasm (n = 3). Scale bars: 50 µm. b qRT-PCR analysis of OGRU expression in the cytoplasmic (Cyt) and nuclear (Nuc) fractions. The control, 45S ribosomal RNA (rRNA), was primarily localized in the nucleus, and 12S rRNA was found primarily in the cytoplasmic fraction (n = 3). c The RegRNA2.0 prediction for the binding sites for candidate miRNAs in the OGRU transcript. d qRT-PCR analysis of miRNA levels after treatment with si-OGRU in MC3T3-E1 cells (n = 3). e qRT-PCR analysis of OGRU expression levels after treatment with mimic-320, inhibitor-320, or the corresponding controls (n = 3). f qRT-PCR measurement of the relative miR-320-3p levels in MC3T3-E1 cells under clinorotation-unloading condition for 24, 48, and 72 h (n = 3). g qRT-PCR measurement of miR-320-3p levels after MC3T3-E1 cells were transfected with pcDNA3.1(+)–OGRU and subjected to clinorotation unloading for 48 h (n = 3). h qRT-PCR detection of the miR-320-3p levels in the tibias from mice in the CON, HLU, HLU + (DSS)₆–liposome and HLU + (DSS)₆–liposome–OGRU groups (n = 5). i, j The relative luciferase activity of luciferase reporters containing OGRU wild-type (WT) or mutated (MUT) miR-320-binding sites in 293T cells cotransfected with mimic-320 or its negative control was measured. The data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity (n = 3). k Anti-AGO2 RIP was performed using input from cell lysate, normal mouse IgG, or anti-Ago2, followed by qRT-PCR to measure the OGRU and miR-320 levels (n = 3). All data are the mean ± SD. *P < 0.05, **P < 0.01.