Figure 6.

Binding of α-catenin to β-catenin and actin is required for JNK-mediated AJ formation. A) Schematics of different domains of α-catenin. ME180 cells were transduced with recombinant lentivirus encoding for different domains of α-catenin as indicated, each fused with HA tag to facilitate detection. B) Cell lysates were subjected to Western blotting for detecting total amount of E-cadherin, β-catenin, and HA-tagged α-catenin domains. C) ME180 cells expressing different domains of α-catenin were treated with either vehicle (ctrl) or SP600125 (10 μM) for 30 min. Cell lysates were immunoprecipitated with antibody against β-catenin and subsequently subjected to Western blotting for HA tag and E-cadherin. β-Catenin was used as loading control. D) Each band intensity in C was normalized to the corresponding β-catenin and plotted as mean ± sd of 3 independent experiments. E, F) ME180 cells expressing α-catenin (E) or α-Δactin (F) were treated with either vehicle or SP600125 (10 μM) for 30 min. Triton-X100 soluble (S) and insoluble (I) fractions were subjected to Western blotting for E-cadherin, β-catenin, and HA tag.