Fig. 2: MiR-23b-3p inhibits HS6ST2 expression by targeting HS6ST2 mRNA and effects specific gene expression in chondrocytes.

a, b Dual luciferase reporter assay to validate target relationship between HS6ST2 and miR-23b-3p. SW1353 cells were transfected with mimic miR-23b-3p (a) or anti-miR-23b-3p (b) by control vector, pmirGLO-HS6ST2 wild-type 3′UTR vector or pmirGLO-HS6ST2 mutant 3′UTR vector, respectively, for 48 h. c, d Stem-loop RT-qPCR and western blotting results in SW1353 cells (c) or C28/I2 cells (d) transfected with 10 nM mimic miR-23b-3p (left panel) or 50 nM anti-miR-23b-3p sequence (right panel). e, f RT-qPCR (e) and western blotting (f) results of cartilage-specific gene expression in SW1353 cells transfected with mimic miR-23b-3p or negative control. g, h RT-qPCR (g) and western blotting (h) results of cartilage-specific gene expression in SW1353 cells transfected with anti-miR-23b-3p or negative control. RNA was harvested at 24 h, while protein was isolated at 48 h after transfection. U6 snRNA was used as internal controls in miRNA stem-loop RT-qPCR detection and GAPDH was used as internal controls in mRNA RT-qPCR and western blotting detection. Bars represent standard error of the mean (SEM) from three independent experiments. Mann–Whitney U test was used to identify statistical differences between two groups. *P value < 0.05