FMNL2 associates with AJ components. (A) E-Cadherin was immunoprecipitated (IP) from cells expressing GFP or GFP-Rac1-L61 to test for coprecipitation of FMNL2 by immunoblotting. (B) Detection of endogenous E-Cadherin in immunoprecipitations of FMNL2 from wild-type MCF10A cells by immunoblotting. (C) HEK cells were transfected with E-Cadherin–GFP and FLAG-tagged FMNL2 derivatives. FLAG-tagged proteins were precipitated and bound GFP-tagged protein was detected by immunoblotting. (D) HEK cells were transfected with FLAG-FMNL2 N terminus (NT) and myc-FMNL2 CT. FLAG-FMNL2 NT was precipitated and tested for coprecipitation of myc-FMNL2 CT in the presence of increasing amounts of E-Cadherin CT. Purified E-Cadherin CT peptide was visualized by Coomassie staining. Numbers indicate statistical values (n = 3, means ± SD) obtained by quantification of coprecipitated myc-FMNL2 CT. (E) Control experiment to C using purified GFP. (F) Experiments were performed as in Fig. 3 B with α-catenin–GFP. (G) GST or GST–α-catenin were purified from bacterial lysates and incubated with MCF10A cell lysate. GST–α-catenin pulled down endogenous FMNL2 as well as β-catenin.