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Dispase lifting resulted in cell sheet contraction, with disruption of actin or intermediate filaments, but not microtubules, significantly reducing the amount of cell sheet contraction.
Blebbistatin treatment significantly reduced cell sheet contraction, and this effect was transient if blebbistatin was removed.
Sheet areas were then used to compare the effect of blebbistatin on sheet contraction.
Blots were developed with ECL reagents (Perkin Elmer) and imaged using a FUJI imaging unit (Fujifilm, Stamford, CT). ANOVA was performed on the cell sheet contraction data.
The relative impact of each component of the cytoskeleton on cell sheet cohesion and cell sheet contraction is not currently well-established.
Timing of TSB synthesis was critical and needed to be performed upon first observation of cell sheet contraction as described above.
Similar(42)
Both passive and active processes are likely involved in sheet contractions.
Whole-sheet contraction was measured without nuclei labeling during blebbistatin experiments, and Microsoft Paint was used to quantify cell sheet diameters and subsequently, sheet areas.
Keratin intermediate filament disruption caused comparable effects on cell sheet cohesion and contraction, when compared to actin or microtubule disruption.
All components of the cytoskeleton are involved in maintaining cell sheet cohesion and contraction, although not to the same extent.
Thus, results from the characterization of how different cytoskeletal components contribute to cell sheet cohesion and contraction have broad application.
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