Figure 3
TGF-β and BMP reporter assay analysis of Smad4 -mutant embryonic stem cell lines. TGF-β and BMP reporter assay analysis of Smad4-mutant embryonic stem (ES) cell lines. A. TGF-β (CAGA12-MLP-luciferase) and B. BMP (BRE-luciferase) reporter assays were carried out in wild-type (+/+), Smad4+/E6sad(+/-), and Smad4E6sad/E6sad(-/-) ES cell lines. Normalized CAGA-luc and BRE-luc levels are indicated for each cell line. For each genotype reporter activities are shown for cells transfected (TBRII+ and ALK2+) and non transfected (TBRII- and ALK2-) with the corresponding receptor-expressing vector. In C. the TGF-β reporter assays analysis was carried out after transfection with a SMAD4-expressing vector. Each bar represents the average of three independent experiments, and the error bars represent the standard deviation. For the CAGA12-MLP-luciferase assay, three independent ES wild-type clones have been employed. The WT bar represents the average of the luciferase activities measured in two independent experiments. p values were calculated by the statistical two samples t-test (two tails).