Figure 2

t-DARPP increases nuclear accumulation of β-catenin. Immunofluorescence analysis was performed on AGS (A) MKN28 (B) and FLO-1 (C) cells overexpressing t-DARPP. Data indicates increased nuclear localization of β-catenin in cells expressing exogenous t-DARPP, as compared to control cells that show membranous β-catenin (p < 0.001). β-catenin positive cells were analyzed by using an anti-β-catenin antibody which is recognized by secondary rabbit antibody conjugated with FITC, depicted by green fluorescence. Nuclear staining was detected by counterstaining cells with 4', 6-Diamidino-2-phenylindole (DAPI), represented as blue fluorescence. Ratio of cells positive for nuclear β-catenin staining to total number of cells was counted as percentage positive for nuclear localization. In total, at least 300 cells from t-DARPP and control vector were counted from three different microscopic fields for β-catenin immunofluorescence. Results are representative of three independent experiments and expressed as mean values ± SD. Significance of difference was calculated using Student's t test.