Figure 5

Fibronectin regulates CCL2, ILK and α5 integrin. HCT116 cells were plated on tissue culture plates coated with increasing levels of fibronectin for 4 h. In A, the CCL2 levels were measured in the medium using ELISA. In B, the cells were subsequently lysed and equivalent amounts of protein resolved using SDS page. After transfer the membranes were probed with the antibodies indicated. The data indicate increased ILK and α5 integrin expression with increasing fibronectin exposure, peaking at 20 ug/ml. C. Immunohistochemistry was performed on colonic sections using a fibronectin antibody on 3 different mice in each group. There is a clear reduction in fibronectin expression in the ILK knockout mice (a representative slide is shown). D. HCT116 cells were exposed to the doses of fibronectin indicated and the QLT 0267 compound added at the start of the experiment. The cell lysates were then resolved using SDS-PAGE and probed with either the α5 integrin or beta actin (loading control) antibodies. The data are representative of 3 separate experiments.