Inhibition of Wnt3a-induced ALP activity by Dkk1 and -2 in cell lines C2C12 and C3H10T1/2

<p><b>Copyright information:</b></p><p>Taken from "Attenuation of WNT signaling by DKK-1 and -2 regulates BMP2-induced osteoblast differentiation and expression of OPG, RANKL and M-CSF"</p><p>http://www.molecular-cancer.com/content/6/1/71</p><p>Molecular Cancer 2007;6():71-71.</p><p>Published online 30 Oct 2007</p><p>PMCID:PMC2173906.</p><p></p> Cells were pre-incubated for 3 h with Dkk1-4-CM or M-CM. Wnt3a-CM was added and cells were cultured for six days. L-CM was used in control experiments to determine the baseline level of ALP in cells not stimulated with Wnt3a (not shown). ALP activity was measured by ELISA and values were normalized to total protein. Results are presented as fold induction relative to cells not stimulated with Wnt3a. Data are means ± SD from three independent experiments. Asterisks indicate significant decreases (< 0.005) compared to cells not treated with Dkk. , Inhibition of Wnt3a-induced ALP mRNA by Dkk1 and -2. Experimental setup was the same as described in panel A except cells were stimulated with Wnt3a for 3 rather than 6 days. ALP mRNA was determined by real-time RT-PCR and normalized to the message levels of β-actin mRNA. Data are means ± SD from three independent experiments. One asterisk (< 0.05) and two asterisks (< 0.005) denote a significant loss of ALP message compared to cells not treated with Dkk. , Inhibition of Wnt3a-induced OCN mRNA expressions by Dkk1 and -2. Experimental setup was the same as described in panel A. OCN mRNA was determined by real-time RT-PCR and normalized to β-actin message levels. Data are means ± SD from three independent experiments. One asterisk (< 0.05) and two asterisks (< 0.005) denote a significant drop in OCN message relative to cells not treated with Dkk