Xenbase Image ID: 135812

Figure 5.  Smad6 interact with itself and Smad1, but not with Smad2 or Smad4. (A) Specific interaction between Smad1 and Smad6. COS cells were transiently transfected with Flag-tagged Smads as indicated on the top, and HA-tagged Smad6 (toppanel) or Smad6 C-domain (bottom panel). Cells were treated with 5 nM BMP2 or 100 pM TGFβ for 1 hr before harvest. Cell lysates were subjected to immunoprecipitation with anti-Flag antibody and then immunoblotting using the anti-HA monoclonal antibody 12CA5. Expression of Smads was measured by anti-Flag or anti-HA immunoprecipitation of aliquots of cell lysates, followed by anti-Flag or anti-HA immunoblotting (bottompanel). [Ig(H) and Ig(L)] Immunoglobulin heavy and light chain bands, respectively. (B) The interaction between Smad6 C-domain and Smad1 C-domain is specific and direct. The C-domains of Smad1, Smad2, Smad4, and Smad6 fused to the GAL4 activation domain (GAD) were tested for interaction with Smad6 C-domain fused to the LexA DNA-binding domain in yeast. Interaction was monitored by the β-galactosidase assay, which allowed us to score for association by the presence of blue color. Image published in: Hata A et al. (1998) Copyright © 1998. Image reproduced on with permission of the Publisher, Cold Spring Harbor Laboratory Press. This is an Open Access article. Larger Image Printer Friendly View

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