Figure 3.

Validation that murine miR-342 targets and controls the expression of Col1a2. A) Bioinformatics prediction of the binding region of mouse miR-342 (mmu-miR-342-3p) within the Col1a2 3′-UTR. B) Sequence comparison of the Col1a2 3′-UTR (WT) and the core sequence DM. These fragments were amplified, isolated, and inserted into the luciferase expression vector, pmirGLO, to examine luciferase reporter activity. C) In HEK293T cells transfected with WT Col1a2, an miR-342 mimic inhibited luciferase activity, and an miR-342 inhibitor enhanced luciferase activity. D) In HEK293 cells transfected with the DM, the miR-342 mimic lost the ability to inhibit luciferase activity. E, F) In osteogenic cells, the miR-342 inhibitor increased Col1a2 RNA (E, left) and protein (E, right) expression, but an miR-342 mimic reduced Col1a2 protein expression (F, right). *P < 0.05, compared to NC. Data are means ± sd.