Matricellular Protein SMOC2 Potentiates BMP9-Induced Osteogenic Differentiation in Mesenchymal Stem Cells through the Enhancement of FAK/PI3K/AKT Signaling

Figure 6

The activation of FAK/AKT signaling may be triggered by facilitating the interaction of SMOC2 and integrin β1 in MEFs. (a) Western blot assay results showed the effect of SMOC2 and/or siFAK on the phosphorylation levels of FAK and AKT promoted by BMP9 in MEFs at 30 h. (b–c) Quantification of the western blot results showed the effect of SMOC2 and/or siFAK on the phosphorylation levels of FAK and AKT promoted by BMP9 in MEFs. (d) ALP assay results showed the effect of LY290042 on the activity of ALP induced by BMP9 in MEFs. (e) ALP assay results showed the effect of SMOC2 and/or LY290042 on BMP9-induced ALP activity in MEFs. (f) Quantification results of the ALP assay showed the effect of LY290042 on the activity of ALP induced by BMP9 in MEFs. (g) Quantification results of the ALP assay showed the effect of SMOC2 and/or LY290042 on BMP9-induced ALP activity in MEFs. (h) Alizarin Red S staining showed the effect of SMOC2 and/or LY290042 on BMP9-induced mineralization in MEFs. (i) Quantification results of Alizarin Red S staining showed the effect of SMOC2 and/or Ly294002 on the effect of BMP9-induced mineralization in MEFs. (j) Confocal laser scanning immunofluorescence staining results showed the effect of SMOC2 on the phosphorylation of AKT induced by BMP9 in MEFs at 30 h. (k) ChIP assay shows the enrichment of GTF2I at the promoter region of smoc2 in MEFs. (l) Immunoprecipitation assay (IP) results showed that SMOC2 may interact with integrin β1 in MEFs. Compared with the control group, ; ; compared with the AdBMP9 group, #, ##; compared with the AdBMP9 and LY290042 group, ^, ^^.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

(k)

(l)