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. 2014 Mar 27;4:4493. doi: 10.1038/srep04493

Figure 5. Wnt5a and BMP2 cooperatively enhance Lrp5 expression through Sp7 in calvarial cells.

Figure 5

(A) Chromatin immunoprecipitation assay of the Lrp5 and Lrp6 genes in wild-type calvarial cells. Calvarial cells were cultured in osteogenic medium for the indicated time and subjected to ChiP analysis. Upper schemas show the promoter of the Lrp 5 and Lrp6 genes. Putative Sp1 binding sites were indicated. E1; exon 1. Asterisks indicate non-specific bands. The full length gels were presented in Supplementary Fig. S5. (B) Effects of Sp7 on Lrp 5 expression in wild-type calvarial cells. After the adenovirus-mediated gene transfer of LacZ (Control) or Sp7 cDNA, cells were cultured in osteogenic medium for 2 days and subjected to Western blot analysis. The full length blots were presented in Supplementary Fig. S5. (C) Effects of Wnt5a and BMP2 on Sp7 expression in Wnt5a−/− calvarial cells. Cells were cultured in the presence or absence of Wnt5a (200 ng/ml) with or without BMP2 (100 ng/ml). The expression of Sp7 mRNA was examined in these cultures. *p < 0.05, **p < 0.01 (D) The expression of Lrp5 and Lrp6 mRNA in Ror2−/− calvarial cells. Wild-type (WT) and Ror2−/− calvarial cells were cultured under osteogenic conditions and subjected to real-time PCR analysis. In (C, D), data are expressed as the mean ± SD (n = 3).