Figure 5

Biological activity of the Dlx3 and Osx in mouse Dspp promoter and protein interactions between Dlx3 and Osx. (A) Illustration of core sequences of Dlx3I, Dlx3II and Osx binding elements. (B) Illustration of wild type and mutant p318 luciferase reporter gene constructs. The mutant region was marked with a cross. (C) The MD10-F2 cells were co-transfected with wild type or mutant p318 luciferase gene constructs with pcDNA-Dlx3 or/and pcDNA-Osx-Flag expression plasmids. Luciferase activity was determined, and the value obtained from the wild type p318 group was taken as 1-fold. *P < 0.05; **P < 0.01; compared with wild type p318 group. (D) Anti-Osx, anti-Dlx3 antibodies and negative control IgG were used to pull down by co-immunoprecipitation assay. Anti-Dlx3 or anti-Osx antibody was used for western blotting to confirm the presence of Dlx3 or Osx in the complex. Neg.: Negative control IgG; W.B.: Western blot. Uncropped images of blots were shown in Supplementary Fig. S5.