Fig. 1.

Effect of silencing β-catenin on mesalamine (5-ASA) induced membranous translocation of E-cadherin. (a) HT29 cells were transfected with control and siβ-catenin RNA. Cells were grown on coverslips and treated with 5-ASA (20 mM) for 24 h. Immunostaining was performed using antibodies against β-catenin and E-cadherin. 5-ASA induced membranous translocation of both proteins. An increased interaction of β-catenin and E-cadherin was observed upon 5-ASA treatment in cells transfected with control RNA (merged image). Membranous turnover of E-cadherin was unaffected upon 5-ASA treatment in siβ-catenin transfected cells. Image magnification 400× (b) Western blot analysis of E-cadherin and β-catenin in total cell lysate (RIPA) and cellular fractions (membrane or cytoplasm) in siβ-catenin transfected cells. Anti-tubulin was used as loading control and Na–K ATPase was used to verify the purity of membranous fraction.