Fig. 4

SMAD7 was a direct target of miR-16. a Sequence alignment of miR-16 with the putative binding sites within in MEG3 and mutant miR-16 binding sites. b Dual-luciferase reporter assays were used to investigate whether SMAD7 could directly interact with miR-16 by the putative binding sites in HEK 293T cells cotransfected with wild-type or mutant-type MEG3 luciferase vectors and miR-NC or miR-16 mimics. c SMAD7 expression was detected in IL-1β-induced chondrocytes transfected with miR-NC, miR-16 mimics, anti-miR-NC or anti-miR-16. β-actin was used as internal reference. d The effect of MEG3 on luciferase activity in HEK 293T cells transfected with SMAD7 (WT) vector and miR-16 mimics was determined. e SMAD7 expression pattern was detected by western blot assay in IL-1β-induced chondrocytes transfected with pcDNA-NC, MEG3, miR-NC or miR-16. f SMAD7 expression pattern was detected in IL-1β-induced chondrocytes transfected with si-NC, si-MEG3, anti-miR-NC or anti-miR-16. *P < 0.05 vs. negative control