Analysis of morphology and migration of fibrosarcoma cells exposed to contractility inhibitors.

<p><b>a</b>): Cells were treated with P6 (5 µM), blebbistatin (5 µM) or Y27632 (10 µM) for 18 h. Representative images of P6 treated cells by phase contrast microscopy; blebbistatin-treated cells stained for F-actin; Y27632 treated cells stained with CellMask Orange. Scale bars, 50 µm. <b>b</b>): Migration was quantified by measuring wound closure. Images were collected at 12 h for the fswt and fs188 cells, at 18 h for the fs120 cells and at 24 h for the fs164 cells to correlate with 40–60% wound closure in untreated conditions. Contractility inhibitors were added at the same concentrations used in (<b>a</b>) 18 h before the start of the assay and replaced in fresh media at assay start. Each point represents data obtained from 3 independent experiments ± SEM, each conducted with 2–4 replicates. *p<0.05, ***p<0.001 values represent significance over untreated cultures (ANOVA followed by Tukey-Kramer post-test). <b>c</b>): Cells were incubated overnight with P6 (5 µM). Equal amounts of proteins (30 µg) were analysed for integrin β1 (β-Itg) and pMLC. A representative immunoblot is shown.