Angiomotin family members directly interact with Patj via different PDZ domains. (A) Domain architecture of Patj constructs. Full-length Patj (1801 aa's) consists of one N-terminal L27 (blue) and 10 PDZ domains (orange). (B) Summary of the Y2H assays. S cerevisiae MaV203 cells were cotransformed with bait constructs containing cDNAs encoding different Patj deletion mutants and prey plasmids that encode the C-termini of Amot, AmotL1, AmotL2, and Syx as well as Syx lacking the last 2 amino acids. To test for interaction in the Y2H assays, corresponding strains were cultivated in synthetic media lacking leucine, tryptophan, and histidine, supplemented with 50 to 75 mM 3AT (his3). In addition, β-galactosidase reporter gene activity (lacZ) was determined on replica filters using X-gal as substrate. − indicates no growth in selection media or no β-galactosidase activity; +/−, background growth or background β-galactosidase activity; +, growth in selection media or weak β-galactosidase activity; +/+, very fast growth in selection media or high β-galactosidase activity; and nd, not determined. (C) Figure summarizing the binding specificities of Amot, AmotL1, AmotL2, and Syx to individual PDZ domains of Patj. The binding of Pals1 to L27 was shown by Roh et al.³⁴