To study the molecular mechanism by which nonmuscle myosin II (MII) regulates protrusion and adhesion dynamics in migrating cells, NIH3T3 cells were treated with βPIX siRNA (Scr) for 2 days, treated with DMSO for 30 min., and stained for βPIX (green) and vinculin (red). In scrambled siRNA-treated cells treated with DMSO, vinculin localized to large focal adhesions at the cell periphery that overlapped with βPIX. βPIX siRNA had little effect on its own. These findings help elucidate a functional link between MII and Rac1/Cdc42 GTPases, which may regulate protrusion/adhesion dynamics in migrating cells. This image is the original data file from Fig. 6A, “Requirement for βPIX in MII-regulated cell protrusion and adhesion.” J. Cell Biol. 2010. Vol. 190(4):663–674.
Cells were cultured in DME (Invitrogen) supplemented with 10% fetal bovine serum and 100 U/ml penicillin/streptomycin (Invitrogen) at 37°C in a humidified 5% CO2 incubator. siRNA transfected NIH3T3 cells were treated with DMSO for 30 min, and co-stained for betaPIX (green) and vinculin (red). Images were captured by Olympus IX81-ZDC inverted microscope with UPlanFL N 40X objective.
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