HMGB1 increases adhesiveness of enterocytes to the underlying matrix in a TLR4-dependent manner. A, single cell force contraction microscopy of IEC-6 cells after transfection with the indicated construct and treatment with PBS or HMGB1 (100 ng/ml); the region of interest is shown in white. Bar = 5 μm. WT, wild type; DN, dominant negative. Quantification is shown in B. *, p < 0.05 versus PBS paired control. Results shown are representative of three separate experiments. C and D, SDS-PAGE showing expression of active and total RhoA (C) or phosphorylated and total cofilin (D) in IEC-6 cells after transfection with indicated construct ± HMGB1 or LPS (50 μg/ml) as indicated. Results shown are representative of three separate experiments. Ctrl, control. E, IEC-6 migration after treatment with HMGB1 ± Rho kinase inhibitor Y27632. Results shown are representative of three separate experiments. *, p < 0.05 versus control. Error bars indicate S.E.