Sulindac Sulfide Effects on HCT15 Cells

HCT15 cells (CCL-225; American Type Culture Collection) were treated with 50 µM sulindac sulfide (Sigma-Aldrich) for 1-24 h and analysed by western blotting following our previously established protocols (Mladenova et al., 2011 (link), 2013 (link)). Only adherent cells were analysed for protein expression. The membranes were incubated with primary antibodies for 1 h at room temperature or overnight at 4°C (β-actin, 1:10,000, clone AC15, Sigma-Aldrich), phosphorylated c-Jun (Ser73) no. 9164, c-Jun no. 9165 (Cell Signaling Technology), E-cadherin (1:2000, no. 610181, BD Biosciences), β-catenin (1:2000, no. 610153, BD Biosciences), p120-catenin (1:1000, clone H-90, no. sc-13957, Santa Cruz Biotechnology). ImageJ densitometry software (National Institutes of Health) was used for quantitative densitometry analysis.