Cd2+ disrupts the adherens junction complex and redistributes β-catenin from the periphery to cytosol and nuclei of kidney PTC. (A) Time dependent increase of β-catenin protein distribution in cytoplasmic and nuclear fractions of WKPT-0293 Cl.2 cells without (ctl) or with Cd2+. Cytosolic and nuclear fractions were immunoblotted with β-catenin antiserum. For loading controls, the same membranes were reprobed with antibodies to β-actin and γ-tubulin for cytoplasmic and nuclear fractions, respectively. β-catenin signals were normalized to loading markers and compared to controls, which were set to 100%. Means ± SEM of 3 experiments are shown. One-way ANOVA was used assuming equality of variance with Levene's test and Tukey post hoc test for pair-wise comparison between control and Cd2+ exposed cells. (B) Immunofluorescence staining patterns of β-catenin in WKPT-0293 Cl.2 cells ± Cd2+. Nuclei were stained with propidium iodide (PI). Note the peripheral β-catenin labeling in control cells whereas in Cd2+ exposed cells β-catenin is found diffusely distributed in cytosol and nuclei. Bars = 20 μm. (C) Distribution of α-catenin in cytoplasmic and nuclear fractions increases with Cd2+ exposure time, as shown by immunoblotting. (D) Cd2+ reduces association of β-catenin with E-cadherin and α-catenin as a function of time. Cell lysates were immunoprecipitated (IP) with β-catenin antiserum and immunoblotted (IB) with E-cadherin or α-catenin antisera.