Figure 4

H. pylori -induced nuclear accumulation of β-catenin, activation of LEF/TCF and target gene expression depends on LRP6, Dvl2 and Dvl3. A NCI-N87 cells, control shRNA, Dvl2 shRNA cells, Dvl3 shRNA cells, and LRP6 shRNA cells were infected for 1 h with H. pylori P1 wildtype strain. Nuclear fractions were prepared (S-PEK Kit, Calbiochem). Western blot analysis was performed with antibodies as indicated. Diagram shows the relative amounts of β-catenin after densitometric analysis from two independent experiments. Densitometric values for β-catenin were normalized to HDAC1 content in each sample and depicted relative to the values of the respective non-infected cells. B, Cells were transiently transfected with 8 × SuperTOPflash and cotransfected with renilla luciferase plasmid. 24 h later cells were infected with H. pylori P1 wildtype strain for the indicated periods of time, lysed and firefly/renilla luciferase activity was measured. Diagram represents the mean fold changes + SEM of 3 separate experiments performed in triplicate. * p < 0.05, relative to activity in NCI-N87 parental cells. C, NCI-N87 cells, control shRNA cells, Dvl2 shRNA cells, Dvl3 shRNA cells, and LRP6 shRNA cells were infected for 3 h with H. pylori P1 wt. Whole cell RIPA lysates were prepared. Western blot analysis was performed with antibodies as indicated. Diagram shows the relative amounts of Axin2. Densitometric values for Axin2 were normalized to GAPDH content in each sample and depicted relative to the values of the respective non-infected cells.