Figure 6.

Treatment of Yoda1 upregulates expression of WNT16 gene which promotes mineralization of SHED. (a,b) Expression of WNT and DACT3 genes. SHED were cultured with or without 5 μM Yoda1 for 24 hrs. Total RNA was prepared from the cells and analyzed by real-time RT-PCR for WNT expression (a) and DACT3 expression (b). (c) Alizarin Red S staining after WNT16 treatment to SHED. After 7 days of culture of SHED in odontogenic induction media in the presence or absence of 300 ng/ml exogenous WNT16, Alizarin Red staining was performed in SHED. The Alizarin Red-positive areas were quantified using ImageJ. Scale bar, 150 μm. The data, as shown, are representative of three independent experiments with similar results, and error bars indicate standard deviations. Statistical analysis was performed using analysis of variance (**p < 0.01).