Interaction of casein kinase 1 epsilon mutants and Dishevelled 2. Casein kinase 1 epsilon (CK1ε) mutants associate with Dvl2, co-localize with Dvl2, and promote the punctate cytoplasmic localization of Dvl2. (a) HEK293 cell lysates transfected with either wild-type (WT) CK1ε or the P3, P4, and P6 mutants together with Dvl2-Myc were lysed and immunoprecipitated using an anti-Myc antibody. WT CK1ε and all of the CK1ε mutants efficiently bind the Dvl2 protein. (b) Dvl2-Myc protein localization in transfected COS7 cells was observed by confocal microscopy using an anti-Myc antibody. Dvl2 is either found in cytoplasmic inclusions or evenly dispersed within the cytoplasm. (c) WT CK1ε co-transfected with Dvl2-Myc into COS7 cells dissolves most of the Dvl2 punctae, resulting in the predominance of evenly distributed Dvl2 protein. In contrast, the P3, P4, and P6 mutants enhance the formation of Dvl2 clusters with a punctate appearance in the cytoplasm. A localization pattern of CK1ε was observed with an anti-CK1e antibody. All CK1ε forms show even cytoplasmic distribution when transfected alone. (d), (e) Cells with an even or punctate distribution of Dvl2 and CK1e were counted. For each condition, at least 150 cells were examined.