FIGURE 3.

A–D, HEK-293 cells were transfected with the indicated constructs and TopFlash reporter. Twenty-four hours post-transfection, cells were treated with CK1 (D4476 100 μm, IC261 50 μm) and CK2 (TBCA 50 μm, TBBt 100 μm) inhibitors. Both CK1 and CK2 inhibitors were able to significantly block TCF/LEF-driven transcription in both FLAG-Dvl3 + CK1ϵ (A)- or FLAG-Dvl3+CK2/PAR1 (B)-transfected cells. The inhibitors did not show any effect in cells transfected with constitutively active β-catenin (C) or in the absence of overexpressed Dvl3 (D). E and F, HEK-293 cells were transfected with siRNA against CK1ϵ or CK2α. After 24 h they were transfected with the corresponding constructs and TopFlash + Renilla constructs. siRNA against CK1ϵ and CK2α were able to diminish Dvl+CK1ϵ-induced TopFlash, whereas Dvl+CK2/PAR1-induced TopFlash could not be diminished by the siRNA. Data represent the means ± S.D. **, p < 0.01; ***, p < 0.001; one-way analysis of variance, Tukey post test. ns, not significant.