Silencing β-catenin diminishes the synergistic osteogenic activity between BMP9 and Wnt3A in iMEF cells.

<p>(<b>A</b>) Wnt3A and/or BMP9-induced early osteogenic marker alkaline phosphatase (ALP) activity is reduced in iMEF-simBC3 cells. Subconfluent iMEF-simBC3 and iMEF-siControl cells were infected with AdWnt3A, AdBMP9, AdGFP, or AdWnt3A+AdBMP9. At day 5 post infection, cells were fixed for ALP histochemical staining assay. Each assay conditions were done in triplicate. Representative results are shown. (<b>B</b>) Wnt3A and/or BMP9-induced ALP activity is decreased in the β-catenin silenced iMEFs. The experiments were set up in a similar fashion to that described in (<b>A</b>). At days 3 and 5, cells were lysed for quantitative ALP activity assays. Basal ALP activities (e.g., GFP groups) were subtracted from all BMP9, Wnt3A, and Wnt3A+BMP9 groups. Each assay conditions were done in triplicate. “**”, <i>p<0.001</i> (iMEF-simBC3 <i>vs</i>. iMEF-siControl).