Figure 5. miR-133b suppresses osteogenic differentiation of AMSCs through RUNX2. (A, B) Plasmids that overexpress or deplete RUNX2 were validated by RT-qPCR (A) and by ELISA (B) for RUNX2. (C–G) AMSCs were transfected with either miR-133b or as-miR-133b, compared to null controls. In another two groups, AMSCs were co-transfected with miR-133b and RUNX2 or with as-miR-133b and shRUNX2. (C, D) RT-qPCR (C) and ELISA (D) for RUNX2. (E, F) Osteogenic differentiation of AMSCs was determined by Von kossa staining, shown by representative images (E) and by quantification (F). (G) RT-qPCR for osteogenesis-associated genes, collagen I (Col I) and osteopontin (OPN). N=5. *p<0.05. Scale bars are 50μm.