Fig. 9

Phosphorylation of S268 and S311 contributes to the activation of the Wnt/b-catenin pathway. a, b: HEK293 cells were transfected by indicated plasmids together with TopFlash and Renilla reporter plasmids. The ability of individual kinases to induce activation of Wnt/β-catenin pathway either alone (a) or in combination with DVL3 (b) was analyzed. Mean, SD and individual data points are indicated. Statistical differences were tested by One-way ANOVA and Tukey’s post test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). c, d: Rescue experiments with DVL3 S268/S311 mutants. WT and D1/2/3 TKO HEK293 T-Rex cells were transfected as indicated and treated according to the scheme with 80 ng/ml recombinant human Wnt3a (rWnt3a). All samples were treated with 0.1 μM LGK974 inhibitor and 250 ng/ml R-SPONDIN1 and analyzed by TopFlash assay (c). Mean, SD and individual data points are indicated. Statistical differences were tested by paired t-test (* p < 0.05, ** p < 0.01). Samples were also used for WB analysis (d)