Regulation of signaling pathways by TAZ in vitro.

<p>(A) C3H10T1/2 cells were transiently cotransfected with the p6OSE2-Luc plasmid and expression vector for TAZ (pEF-Flag-TAZ) or with the empty vector (pEF-BOS). The cells were also transfected with the expression vector for til (pcDNA3-til) or empty vector as indicated. (B) C3H10T1/2 cells transiently cotransfected with PPRE-Luc and the expression vector for TAZ (pEF-Flag-TAZ) or with the empty vector (pEF-BOS). The cells were also transfected the expression vector for PPARγ (pSPORT-PPARγ) or empty vector as indicated and were exposed to either the vehicle or 1 µM rosiglitazone. Forty-eight hours after transfection, the cells were harvested and used for luminometry. Luciferase activities were normalized for transfection efficiency using <i>renilla</i> activity and all values were expressed as a fold induction relative to basal promoter activity. The results shown are representative of three independent experiments. (C) Primary calvarial cells from Col.1-TAZ (T) or WT (W) mice were treated with 150 ng/ml rhBMP-2 for 2 days and harvested for chromatin immunoprecipitation assays. Soluble chromatins were prepared and precipitated using control IgG or anti-Flag antibodies. The Runx2 binding site in the mouse osteocalcin promoter was amplified by PCR. (D–F) C3H10T1/2 cells were transiently cotransfected with BRE-Luc, p3TP-Lux, and TOPflash and with the expression vector for TAZ (pEF-Flag-TAZ) or an empty vector (pEF-BOS). Cells were exposed to the vehicle or BMP-2, TGF-β, or Wnt-3A CM for BRE-Luc, p3TP-Lux, and TOPflash, respectively. Luciferase activity was measured as described above. Data are expressed as means ± S.D., and the graphical results are representative of three independent experiments, each performed in triplicate (*<i>p</i><0.05 vs. control).