Figure 5
Overexpression and silencing of Mage-D1 affects the mineralization regulation of E19.5 d EMSCs. (a) Under induction with mineralized culture medium for 7 d, the expression levels of Alp, Runx2, Col-1, Dmp1, Dspp were examined by real-time PCR normalized to GAPDH. For raw data see Supplementary Table 3. (b,c) Under induction with mineralized culture medium for 14 d, the expression levels of Opn, Runx2, Ibsp, Dmp1, Dspp were detected by Western blot analysis, GAPDH used as the reference gene. Full-length blots are presented in Supplementary Fig. S4. (d) Under induction with mineralized culture medium for 14 d, Alp staining was used to detect their potential of differential mineralization. Scale bar represents 50 μm. (e) Under induction with mineralized culture medium for 21 d, alizarin red staining was used to detect their mineralized nodules. Scale bar represents 200 μm. (**P < 0.01, ***P < 0.001).
