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Fig. S6 Mapping of Smad3-Binding Domain of Tob1a (A) Reciprocal coimmunoprecipitation between Flag-Smad3 and different Tob1a deletion mutants with HA tag was done in HEK293T cells. Note that Tob1a mutants ND5, MD2 or MD4 failed to coimmunoprecipitate with Flag-Smad3, suggesting that the Box A domain of Tob1a is required for binding Smad3. Schematic draw of Tob1a deletion constructs was shown in Figure 3F. (B) β-Catenin competes with Smad3 for binding Tob1a. Myc-β-Catenin (0, 1, 2 and 5 μg, respectively), Flag-Smad3 (1 μg) and HA-Tob1a (1 μg) were co-transfected into HEK293T cells. Flag-Smad3/HA-Tob1a complexes were immunoprecipitated with anti-Flag antibody and immunoblotted with anti-HA antibody.
Reprinted from Developmental Cell, 11(2), Xiong, B., Rui, Y., Zhang, M., Shi, K., Jia, S., Tian, T., Yin, K., Huang, H., Lin, S., Zhao, X., Chen, Y., Chen, Y.G., Lin, S.C., and Meng, A., Tob1 controls dorsal development of zebrafish embryos by antagonizing maternal beta-catenin transcriptional activity, 225-238, Copyright (2006) with permission from Elsevier. Full text @ Dev. Cell