To study the molecular mechanism by which nonmuscle myosin II (MII) regulates protrusion and adhesion dynamics in migrating cells, Swiss3T3 cells were transfected with a plasmid encoding myc-tagged βPIX (green, stained for myc-βPIX) and costained myosin IIB (red). 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. S1, 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. For transfections, cells in 60-mm-diameter dishes or on fibronectin-coated coverslips were incu- bated with a mixture of DNA and LipofectAMINE 2000 (Invitrogen) according to the manufacturer’s instructions. The cDNA for βPIX was subcloned into pCMV-myc (Takara Bio Inc.). After 4 h, cells were replated onto fibronectin-coated coverslips for 12h and stained for myc- βPIX (green) and MIIB (red). Images were captured by Zeiss LSM 710 confocal microscope with Plan-Apochromat 63X objective.
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