Figure 6.

Direct regulation of RUNX2 by miR-320c. (a) Schematic presentation showing the alignment of miR-320c mature sequence and the putative binding sites within the 3′UTR region of the RUNX2 mRNA using TargetScan database. The exact positions of the interaction between RUNX2 3′UTR and miR-320c seed region are indicated. (b) Overexpression of miR-320c was associated with significant decrease in RUNX2 mRNA and protein. *P<0.05. (c) An illustration of the construction of luciferase reporter vector carrying the predicted RUNX2-miR-320c binding sites downstream of the firefly luciferase gene in the pMIR-REPORT vector. The number of predicted miR-320c binding sites within the 3′UTR region of RUNX2 is shown as black bars. (d) The indicated wild-type or mutant reporter vector was co-transfected with a pre-miR control (100 nM) or pre-miR-320c (100 nM) in HEK-293 cells, and luciferase activity was measured 24 h following transfection. Renilla luciferase activity was used for normalization. Data are presented as mean±S.E, n=6. **P<0.005. (e) A working model depicting the possible mechanisms by which miR-320c promotes adipocytic differentiation of hMSCs through targeting genes involved in multiple genetic pathways