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. Author manuscript; available in PMC: 2017 Jan 19.
Published in final edited form as: Oncogene. 2016 Jun 20;36(3):429–438. doi: 10.1038/onc.2016.209

Figure 3.

Figure 3

PR61ε is required for Wnt signaling responses and binds to Wnt receptors. (a) PR61ε-depleted HEK293 cells (shRNA PR61ε #2558) were transfected with PR61ε-Flag when indicated and stimulated with control or Wnt3a-conditioned medium for 15 min. Total cell extracts were immunoprecipitated with anti-LRP5/6 antibody and associated proteins were analyzed by WB. The arrowhead indicates the migration of phosphorylated Dvl-2 and the asterisk indicates the IgG that hinders the visualization of PR61ε in the immunoprecipitates. (b) HEK293 cells were stimulated with control, Wnt3a or Wnt3a and DKK1-conditioned medium for 30 min. PR61ε was immunoprecipitated from total cell extracts and protein complexes were analyzed by WB. (c) HEK293 cells were treated with control or Wnt3a-conditioned medium for the indicated times. Total cell extracts were immunoprecipitated with anti-PR61ε antibody and associated proteins were analyzed by WB. The quantification of LRP5/6 and Fz proteins bound to PR61ε is also represented. *P<0.05. (d) PR61ε-Flag-overexpressing cells were stimulated with control or Wnt3a-conditioned medium for 15 min. Total cell extracts were immunoprecipitated with anti-Fz antibody and associated proteins were analyzed by WB.