Figure 6.
No change in Met/β-catenin association or tyrosine phosphorylation of β-catenin following HGF treatment in Y654/670F-transfected. Total cell lysates from WT or double mutant-transfected cells cultured with or without HGF were immunoprecipitated with anti-FLAG antibody and blotted for Met, E-cadherin or PY20.
A. A representative western blot analysis reveals decrease in Met-β-catenin coprecipitation in WT-, Y142/654F- and Y142/670F-transfected cells in response to HGF, while no change was evident in Y654/670F-cells. A less dramatic difference was observed in E-cadherin-β-catenin association in all cells. Also phosphotyrosine antibody detected β-catenin tyrosine phosphorylation after HGF treatment in WT-, Y142/654F, Y142/670F and not in Y654/670F cells. Comparative pull down of FLAG-β-catenin in all conditions was confirmed by western with anti-FLAG antibody.
B. A representative analysis of studies done in triplicates demonstrates a decrease in Met-β-catenin association ranging from 3–5 fold in all cells except Y654/670F-transfected cells.
C. A small but consistent decrease in E-cadherin-β-catenin association ranging from 15–30% was observed following HGF treatment in all cells.
D. A 2.5–4.0-fold increase in PY20-FLAG-β-catenin was observed in response to HGF in all cells except Y654/670F-transfected cells.