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CIL:26558*  Cite 

To study the molecular mechanism by which nonmuscle myosin II (MII) regulates protrusion and adhesion dynamics in migrating cells, NIH3T3 cells were treated with 50 µm blebbistatin (BBS) for 15 min. and stained for βPIX (green) and myosin IIB (red). βPIX staining is transformed into numerous small puncta along the membrane after BBS treatment, and that myosin staining is excluded from the leading edge. 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. S4, “Potential involvement of βPIX in blebbistatin-induced morphological changes in MII-regulated cell protrusion and adhesion.” J. Cell Biol. 2010. Vol. 190(4):663–674.

Technical Details

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. Cells were treated with 50 µM blebbistatin, 15min, fixed using 3.7% paraformaldehyde in PBS for 15 min, permeabilized using 0.2% Triton X-100 in PBS for 2 min, blocked with 2% BSA in PBS, and stained with the indicated primary antibodies for at 4°C overnight, followed by incubation with a secondary Alexa Fluor 488–, 546– or 594–conjugated antibody. F-actin was visualized with TRITC or Alexa Fluor 350–conjugated phalloidin. Images were captured by Leica TCS SP2 confocal microscope with HCX PL APO 63X objective.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
Cell Line
Cellular Component
myosin II complex
actin filament
Biological Context
Biological Process
regulation of cell migration
Chan-Soo Lee
Chang-Ki Choi
Eun-Young Shin
Martin Alexander Schwartz
Eung-Gook Kim
J. Cell Biol. 2010. Vol. 190(4):663–674
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
Visualization Methods
Alexa Fluor 488
Alexa Fluor 546
Data Qualifiers
raw, unprocessed data
Sample Preparation
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Spatial Axis Image Size Pixel Size
X 512px ——
Y 512px ——