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Figure 5 | BMC Cell Biology

Figure 5

From: Direct interaction between Smad3, APC10, CDH1 and HEF1 in proteasomal degradation of HEF1

Figure 5

TGF-β type I receptor activation enhances Smad3 interaction with APC10 but does not alter CDH1 interaction with HEF1. (A) Smad3 interaction with APC10 is positively regulated by the activation of TGF-β type I receptor. Flag-Smad3 and T7-APC10 were co-expressed in the presence or absence of a constitutively active TGF-β type I receptor mutant R4T204D (R4TD). The interaction between Smad3 and APC10 was detected by immunoprecipitation of Smad3 with an anti-Smad3 polyclonal antibody followed by Western blot with anti-T7 (Top panel, lanes 4–6). The immunoprecipitated Flag-Smad3 was detected by anti-Flag antibody (middle panel). The expression of R4TD was detected by anti-TGF-β RI polyclonal antibody from Santa Cruz (Bottom panel). R4TD-p represents a potential cleavage product of R4. (B) HEF1 interaction with CDH1 Is not regulated by TGF-β type I receptor activation. The 293 cells were transiently transfected with myc-CDH1 and HEF1 in the presence or absence of Smad3 and R4TD, as indicated. The interaction between HEF1 and CDH1 was detected by immunoprecipitation with anti-p130Cas followed by Western blot with anti-myc (Top panel, lanes 7–13). The expression of myc-CDH1 and Flag-Smad3 was detected by Western blot by anti-myc and anti-Flag (Top panel, lanes 1–6). The expression levels of HEF1 was detected by anti-HEF1 (middle panel) and the levels of R4TD detected by Western blot with anti-TGF-β RI antibody (bottom panel).

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