Fig. 4

miR-30a-5p directly targets Runx2 in MC3T3-E1 cell line. A The sequences of miR-30a-5p binding site within the 3’UTR of Runx2 mRNA predicted by TargetScan, and the constructs of pmir-GLO-Runx2-WT luciferase reporter and pmir-GLO-Runx2-MUT control. B The relative luciferase activity in 293T cells after 24 h co-transfected with miR-30a-5p mimic (miR-30a-5p) or miR-NC and WT or MUT luciferase reporter was determined by dual-luciferase reporter assays. C qRT-PCR measurement of the Runx2 mRNA expressions after 24 h in MC3T3-E1 cells transfected with miR-30a-5p mimic (mimic) or miR-NC and miR-30a-5p inhibitor (inhibitor) or inhibitor NC. D, E RIP assay confirmed the binding between Runx2 and miR-30a-5p in MC3T3-E1 cell line. F RIP assay confirmed that there exists no binding between ALP miR-30a-5p in MC3T3-E1 cell line. G RIP assay confirmed that there exists no binding between OC between miR-30a-5p in MC3T3-E1 cell line. H RIP assay confirmed that there exists no binding between Col1 between miR-30a-5p in MC3T3-E1 cell line. All data are shown as mean ± SEM. *p < 0.05; **p < 0.01