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Figure 3 | Molecular Cancer

Figure 3

From: Attenuation of WNT signaling by DKK-1 and -2 regulates BMP2-induced osteoblast differentiation and expression of OPG, RANKL and M-CSF

Figure 3

Wnt3a inhibits ALP and Osterix, but restores p53, in BMP2-stimulated C2C12 cells. A, Wnt3a diminishes BMP2-dependent expression of ALP. Cells were pre-cultured for 3 h with M-CM (not containing Wnt or BMP inhibitor), Dkk1-CM, or M-CM supplemented with 100 ng/ml Noggin as indicated. L-CM (no Wnt3a) supplemented with 100 ng/ml BMP2 (white columns) or Wnt3a-CM supplemented with the same amount of BMP2 (black bar) was added and cells were cultured for 3 days in case of mRNA measurement or 6 days in case of protein determination. Assays were performed as described in Figure 2. Results are presented as fold induction relative to cells not stimulated with BMP2 or BMP2/Wnt3a. Data are means ± SD from three independent experiments. Asterisks and number signs indicate significant changes (p < 0.005) compared to treatment with BMP2 only (top bar) and BMP2 plus Wnt3a (2nd bar from top), respectively. B, Wnt3a inhibits BMP2-induced expression of Osterix but leaves Cbfa1 unchanged. Message levels of both genes were determined by real-time RT-PCR. Cells were treated and results are presented as described in panel A. C, Wnt3a mitigates the BMP2-dependent loss of p53. Cells were cultured for 3 days as indicated in panel A. Protein levels of p53 (top), p21 (center) and β-actin (bottom) were determined by Western blotting using 20 μg cell lysate per lane. The same result was obtained in two additional experiments (not shown).

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