JCS -- Bryja et al. 120 (4): 586 Figure IG4

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Fig. 4. CK1 ${epsilon}$ is activated by Wnt-5a and binds Dvl2 in dopaminergic cells. (A) SN4741 cell lysates were immunoprecipitated with a control IgG or a CK1 ${epsilon}$ -specific antibody and endogenous or overexpressed CK1 ${epsilon}$ and detected by western blotting. (B) Wnt-3a and Wnt-5a increased the phosphorylation of MBP as detected by western blotting using a specific antibody against phosphorylated serine. Cell lysates from control (1% CHAPS), Wnt-3a or Wnt-5a treated (100 ng/ml) SN4741 cells were immunoprecipitated with a CK1 ${epsilon}$ -specific antibody and subjected to kinase assay using MBP as a substrate. (C) Dvl2-Myc interacts with either CK1 ${epsilon}$ (lane 2) or a kinase-dead CK1 ${epsilon}$ (K>R) (lane 3) mutant in SN4741 cells, as assessed by immunoprecipitation and western blotting with Myc- and CK1 ${epsilon}$ -specific antibodies. Co-precipiated CK1 ${epsilon}$ by the Myc antibody is indicated by ${blacktriangleleft}$ . For all other panels: ${triangleleft}$ , phosphorylated Dvl2-Myc; ${blacktriangleleft}$ , unphosphorylated Dvl2-Myc. (D) Dvl2-Myc interacts with endogenous CK1 ${epsilon}$ as assessed by immunoprecipitation of SN4741 cells transfected with Dvl2-Myc only, by using Myc antibody-conjugated agarose beads (control, G-protein-conjugated beads). Data are representative of three independent replicates.