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

Figure 2. Wnt5a-deficient calvarial cells exhibit impaired osteoblastogenesis associated with a reduction in Lrp5 and Lrp6 expression.

Figure 2

(A) The expression of Lrp5 and Lrp6 in wild-type (WT) and Wnt5a−/− calvarial cells. Calvarial cells were cultured in osteogenic medium and subjected to real-time PCR analysis. (B) Western blot analysis of Lrp5/6 expression in WT and Wnt5a−/− calvarial cells. Cells were cultured using the same method as that described in (A). The full length blots were presented in Supplementary Fig. S5. (C) Cytochemical staining for alkaline phosphatase activity (top. blue) and alizarin red S staining (bottom, red) in WT and Wnt5a−/− calvarial cell cultures in osteogenic medium in the presence of BMP2 (200 ng/ml). (D, E) The expression of osteoblastic transcription factors (D) and marker genes (E) in WT and Wnt5a−/− calvarial cell cultures. Cells were cultured using the same method as that described in (A). In (A, C–E), data are expressed as the mean ± SD (n = 3). *p < 0.05, **p < 0.01.