Fig 1: All Dacts form robust complexes with CK1δ/ε, PKA and PKC; less so with other serine/threonine kinases. In this and all subsequent figures, lysates from cells transiently transfected with plasmids expressing N-terminal FLAG-tagged murine Dact proteins or no Dact (negative control) plus the indicated interactor, underwent coIP using anti-FLAG antibody, were separated by SDS-PAGE, and proteins visualized by immunoblotting with the indicated antibody. All cells are HEK293 unless indicated as HEK293T. Unless otherwise indicated all proteins are murine (or identical to murine). Top = coIP immunoblot, bottom = lysate immunoblots (confirming appropriate protein expression), lanes as marked: 1 = Dact1, 2 = Dact2, 3 = Dact3,-= negative control. A, CK1δ. B, D. melanogaster CK1δ/ε (dbt/dco). C, PKA. D, PKCγ. E, CK2a1. F, CK2a2. G, CK2b. H, GSK3β. I, GSK3α (human).