Figure 3.
Validation of miR-27b binding with the putative binding site in the 3′-UTR of MMP-13 mRNA. A, Luciferase activity in HeLa cells transfected with pMIR-REPORT-Luc-MMP13, pMIR-REPORT-Luc-MMP13 mutant, a negative control (NC) miRNA, and miR-27b mimic. B, Relative expression level of miR-27b in human OA chondrocytes (n = 3) transfected with miR-27b mimic or miR-27b inhibitor and then stimulated or not stimulated with IL-1β for 24 hours. C, Expression of MMP-13 protein in culture supernatants after 24 hours of IL-1β stimulation, as estimated by enzyme-linked immunosorbent assay. Chondrocytes were transfected with pre–miR-27b or miR-2b inhibitor. Bars show the mean and SEM results of 3 independent experiments, each of which was run in triplicate. * = P < 0.05 versus control; # = P < 0.05 versus stimulation with IL-1β only; ⊙ = P > 0.05 versus control. See Figure 1 for other definitions.