Figure 5. MMP9 overexpression is associated with decreased paracellular permeability.
In vitro model of stably transfected human intestinal epithelial cells CaCo2BBE overexpressing MMP9 were used to assess paracellular permeability at 0.5 hour, 1 hour, 2 hours and 4 hours by using 4kD and 40kD FITC dextran molecule. (A) Flux of 4kD FITC ng/ml/min. (B) Flux of 40kD FITC ng/ml/min. Each bar represents mean ± S.E., *p< 0.05, **p< 0.005, ***p< 0.0005. The organization of TJs was assessed by performing WBs (30μg/lane) with whole cell lysates probed with (C i-iii) Claudin-2, Claudin-4 and Occludin respectively. The loading control for each blot was GAPDH. Each blot was a representation of three individual experiments. Densitometry evaluations of the WB is represented by the adjacent bar graph and each bar represents mean ± S.E., *p< 0.05. The integrity of TJs were studied by performing WBs (10μg/lane) for Triton X-100 soluble versus Triton X-100 insoluble fraction (D i-iii) Claudin-2, Occludin and Claudin-4 respectively.