Figure 5

Subtotal inhibition of myosin light chain activation increases migration of cancer cells. (A) MDA-MB-231 breast cancer cells were grown in complete medium, quiesced for 24 hours in serum deprived medium (with 0.5% dialyzed FBS) and incubated with varying concentrations of MLCKinase inhibitor, ML-7. Cells were lysed and immunoblotting of lysates was carried out using SDS-PAGE to detect activated levels of MLC. Shown are one of three similar blots.(B) MDA-MB-231 cells were grown in complete medium until they formed a confluent monolayer. The medium was then replaced by 0.5% dialyzed FBS containing quiescent medium for 24 hours. The monolayer was scraped using a sterile pipet tip, washed three times with PBS and migration of cells in the denuded area was assessed over a period of 24 hours in the presence of increasing doses of MLCKinase inhibitor, ML-7. Shown are mean ± SEM of three experiments each performed in triplicate. In comparison to no ML-7 treatment, P < 0.05 for 3 and 15 μM ML-7 treatments.