Figure 5. Zyxin forms a ternary complex with Siah2 and Lats2 and enhances Siah2–Lats2 binding.

(a,b) Double coimmunoprecipitation was performed as depicted (a) and the interactions were determined by western blotting (b). (c,d) Enhanced interaction between exogenously expressed Myc-Lats2 with Flag-Siah2^RM in the presence of increased amount of ectopic expression of Zyxin (c) and quantification of Myc-Lats2 (normalized to Flag-Siah2^RM) protein levels (d). Ratio t-test was applied. (e) Weakened interaction between exogenously expressed Myc-Lats2 with Flag-Siah2^RM in Zyxin-knockdown HEK293T cells. (f,g) MDA-MB-231 cells were treated with 5 μM MG132 and 5 ng ml⁻¹ TGF-β for 12 h. Endogenous interaction between Lats2 and Siah2 was analysed by coimmunoprecipitation (f) and quantification of Siah2 (normalized to Lats2) protein levels (g). Ratio t-test was applied. All data are mean of n=3 independent experiments. All error bars indicate s.d. *P<0.05.