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. 2018 Mar 14;12(5):611–629. doi: 10.1002/1878-0261.12184

Figure 2.

Figure 2

p120‐catenin, CK1ε, and PR61ε are necessary for the activation of noncanonical Wnt pathway. (A) Control or p120‐catenin‐depleted HEK293T cells were treated with control or Wnt5a‐conditioned medium for 15 min. CK1ε was immunoprecipitated from total cell extracts, and the immunocomplex was incubated with 2 pmol of recombinant GST–p120‐catenin in CK1 phosphorylation conditions. Phosphorylation of Ser268 in GST–p120‐catenin was analyzed by WB with a specific PSer268 p120‐catenin antibody. Signal was densitometered, normalized with respect to the GST–p120‐catenin, and represented. The quantification of three different experiments is shown (mean ± SD). The extent of p120‐catenin downregulation by the shRNA is shown in the bottom panel. (B) Fz2 was immunoprecipitated from control, p120‐catenin, or CK1ε HEK293T CRISPR whole‐cell extracts treated with control or Wnt5a‐conditioned medium for 5 min. Protein complexes were analyzed by WB with the indicated antibodies. (C) HEK293T cells depleted of PR61ε using specific shRNA, or a scrambled shRNA as control, were treated with control or Wnt5a‐conditioned medium for 5 min. Fz2 was immunoprecipitated from total cell extracts, and the protein complex was analyzed by WB.