Inhibition of nmMLCK prevents β-catenin translocation to the nucleus. (A) Caco2 cells were treated with IL-1β for 0, 1.5, 3, or 8 hours followed by nuclear fractionation and β-catenin detection by Western blot. Lamin A/C was probed as loading control. (B) Caco2 cells were treated with vehicle control, ML-7, IL-1β, or ML-7 and IL-1β for 3 hours followed by β-catenin detection in the nuclear fraction by Western blot. Results represent results from 4 separate experiments. (C) Confluent Caco2 cells were treated with vehicle control (PBS + 0.1%BSA), IL-1β (100 ng/mL), or IL-1β and ML-7 (10 μM) for 3 hours then subject to nuclear β-catenin (green) detection by immunocytochemistry analysis. Quantitation represents pixel intensity of β-catenin signal detected in nuclear regions. Results are representative of 3 individual experiments. * p<0.05 relative to vehicle control, ¥ p<0.05 relative to IL-1β alone.