Fig. 6
Mynn associates with Smad1
(A) Flag-tagged Mynn interacts with overexpressed Smad1. HEK293T cells were transfected with plasmids expressing Flag-Mynn and Myc-Smad1, and then harvested for immunoprecipitation with anti-Flag antibody.
(B) Overexpressed Smad1 interacts with endogenous Mynn. HEK293T cells transfected with Myc-tagged Smad1 were subjected to immunoprecipitation with anti-Myc antibody.
(C and D) Detection of Mynn-Smad1 interaction in living cells by BiFC assays. YN-Mynn and YC-Smad1 were transfected alone or simultaneously into HeLa cells. The expression of YN-Mynn or YC-Smad1 was detected by immunostaining with anti-GFP antibody (C). The reconstituted YFP fluorescence was detected by confocal microscopy at 488 nm (D). Scale bars: 10 μm.
(E) BMP4 treatment enhanced the interaction between Mynn and Smad1. HEK293T cells transfected with Myc-tagged Smad1 and Flag-tagged Mynn, and then treated with or without BMP4 for 4 h before being harvested for immunoprecipitation.
(F) Schematic representation of Smad1 truncations.
(G) Mynn binds to the MH2 domain of Smad1.