Fig. 7.

DKK3 potentiates canonical Wnt signalling and affects YAP/β-catenin degradation. a Western blots show β-catenin, TAZ, YAP and tubulin expression in murine WT and KO.9 CAFs after treatment with 100 μg mL⁻¹ cycloheximide (CHX) for the indicated times (in min). Graph represents quantification of the indicated blots (normalised to tubulin) at 180 min relative to the amount at 0 min. b Low and high magnification images of LRP6 (green), F-actin (red) and DAPI (blue) staining of murine WT, KO.9 and KO.9-REC CAFs in 1% FBS. Scale bars, 15 μm. c Images show YAP (green) and DAPI (red) staining of CAF1 after transfection with control (siCtr) and LRP5&6 siRNA (smart-pool). Bar, 50 µm. Graph shows quantification of nuclear relative to cytosolic fluorescent intensity (log₁₀ ratios) of YAP. Lines represent mean ± SEM. d Graph shows gel contraction index of murine WT and KO.9 CAFs after transfection with control (siCtr) or Lrp&6 (siLrp5/6) siRNAs. Bars represent mean ± SEM (n = 3 or more). e Graph shows the area of colonies formed by TS1 MOT co-culture with WT and KO.9 CAFs after transfection with control (siCtr) or Lrp5&6 (siLrp5/6) siRNAs. Bars represent mean ± SEM (n = 584 or more colonies from at least 3 independent experiments). f Graph shows gel contraction of WT and KO CAFs cultured in 5% FBS and stimulated with vehicle or 200 ng mL⁻¹ of Wnt3a or Wnt5a. Bars represent mean ± SEM (n = 3). KO data are merged results of KO.2, KO.7 and KO.9 on triplicate (n = 9). g Western blot showing levels of non-phospho (active) β-catenin (Ser33/37/Thr41), β-catenin, YAP, TAZ and Gapdh in WT and KO.9 CAFs after stimulation with vehicle (−) or 200 ng mL⁻¹ of Wnt3a for 3 h. Graph represents quantification of blots indicating the fold levels normalised to Gapdh. h Images show YAP (green) and DAPI (red) staining of murine WT and KO.9 CAFs after 16 h starvation followed by stimulation with vehicle (Starvation) or 200 ng mL⁻¹ of Wnt3a for 3 h. Bar, 50 µm. Graph shows quantification of nuclear relative to cytosolic fluorescent intensity (log₁₀ ratios) of YAP. Lines represent mean ± SEM. Individual p values for different comparisons are shown. For all graphs, *P < 0.05; **P < 0.01; ***P < 0.001; #P < 0.0001; n.s., non-significant