Wnt5a overexpression and dental follicle stem/progenitor cell proliferation and migration. (A) Transfection of lentiviral green fluorescence protein (GFP)-Wnt5a in dental follicle stem/progenitor cells. (B) GFP+ (Wnt5a+) cells were sorted by fluorescence-activated cell sorting (FACS), yielding >96% cells with positive GFP signal. (C) Real-time quantitative PCR (Taqman) showing Wnt5a overexpression (n = 3; P <0.001). (D) Cell Counting Kit 8 assay was applied to evaluate the influence of Wnt5a on cell proliferation. Proliferation rates of dental follicle stem/progenitor cells were attenuated upon Wnt5a overexpression (n = 3; *P <0.05). (E, F) Wnt5a-free, vector control dental follicle cells migrated through Transwell pores. (G, H) Wnt5a-overexpressed dental follicle cells showing robust migration through Transwell pores. (I) Quantitatively, ~3× more Wnt5a-overexpressed cells migrated than the control (n = 3; **P <0.01). (J) Native dental follicle cells with enhanced Wnt5a expression when exposed to osteogenesis induction medium (OM) for 7, 14, 21 and 28 days by western blot. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.