Skip to main content
. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Cell Signal. 2009 Jun;21(6):916. doi: 10.1016/j.cellsig.2009.02.001

Figure 1. Rspo2/C78Y, Rspo2/C113R and Rspo2/Q70R mutants had markedly reduced activity in Wnt/β-catenin pathway.

Figure 1

(A) Luciferase reporter assay. STF cells were transfected with 6 μg of empty vector or indicated Rspo2 construct and lysed 48 h later for luminescence measurements. Values were normalized according to total protein concentration of cell lysates, and luciferase activity was expressed as fold stimulation relative to results with the empty vector. Lysates were assayed in triplicate; error bars (often not visible in figure) represent S.D. (B) Immunoblot analysis of Rspo2 derivatives, β-catenin and LRP6 in transfected cells. After 48 h, lysates were prepared from transfected cells and blotted for β-catenin, phospho-LRP6, total LRP6, Rspo2 derivatives and HSP70 as a loading control. Free β-catenin was detected after pull-down with GST-E-cadherin. Rspo2 proteins also were visualized in concentrated CM. Note that the entire CM from transfected monolayer cultures were loaded on the gel, whereas only 1/15 of cell lysates were subjected to western blotting.