Ajuba contains functional 14-3-3 binding motifs and selectively interacts with 14-3-3 isoforms. (A) Ajuba contains putative 14-3-3 binding motifs in the preLIM region: Aj-A and Aj-B. The numbers correspond to the locations of the core serine residues within 14-3-3 binding motifs of Ajuba and the alanine substitution mutations. (B) Ajuba interacts with 14-3-3 isoforms γ, η and τ when co-expressed in 293 cells. The “*” indicates a non-specific band. (C) Aj-B is the dominant site for 14-3-3 binding. Myc-Ajuba and its mutants, together with Flag-14-3-3γ were co-expressed in 293 cells and co-IP assays were performed. (D) Snail, Ajuba and 14-3-3 can form a ternary protein complex in living cells. Myc-Ajuba, Myc-ΔAj-AB, HA-Snail, HA-ΔSNA-AB and/or Flag-14-3-3γ were expressed in 293 cells, and co-IP assays were performed with α-Myc antibody and western blot with α-HA or α-Flag antibodies.