Figure 3.

c-Met kinase inhibitor (SU11274) inhibits MUC1-dependent in vitro wound healing. MDA-MB-231-control and MDA-MB-231-MUC1 cells (A–F) or BT20 breast cancer cells (I–N) were plated and scratched as described in Figure 1 on both Collagen IV and Fibronectin substrates, after which cells were treated with either DMSO vehicle (A and B), complete media (I and J), 10 ng/mL EGF (A–D and K–L), or 10 ng/mL EGF plus 5 μM SU11274 (E–F and M–N), a selective c-Met kinase inhibitor. Images shown are on Collagen IV substrate. Area filled was determined as in Figure 1. Quantification of area filled is shown in G and H (MDA-MB-231) and O (BT20). (P) Protein lysates were created from BT20 cells grown on Fibronectin overnight in either complete media (left lane), complete media plus 10 ng/mL EGF (middle lane), or complete media plus 10 ng/mL EGF and 5 μM SU11274 (right lane). 40 μg of lysate per treatment were separated by SDS-PAGE and probed by immunoblot to evaluate c-Met phosphorylation (Cell Signaling 3D7), total c-Met (Santa Cruz C-28), and MUC1 expression (NeoMarkers AB-5). Scale bar represents 100 μm. * p<0.05, ** p<0.01, *** p<0.001.