Figure 4.

Wnt3a-mediated phosphorylation of LRP6 is inhibited by Merlin. (a) HEK293T cells were transfected with Flag-Merlin and treated with Wnt3a-CM for the indicated time before being harvested and subjected to SDS-PAGE, followed by western blotting with the indicated antibodies. (b) siGFP- or siMerlin-transfected HEK293T cells were treated with or without Wnt3a-CM for 60 min. Each sample was lysed, subjected to SDS-PAGE, and immunoblotted with the indicated antibodies. The relative level of phopho-LRP6 using the Image J program was shown below the phosphor-LRP6 blot. (c) Plasmids encoding VSVG-LRP6, Myc-Axin, and HA-GSK3β were transfected with or without Flag-Merlin into HEK293T cells. After lysis, each sample was immunoprecipitated with anti-VSVG antibody and immunoblotted with the indicated antibodies. All western blotting and immunoprecipitation experiments were performed more than three times, and these data are representative