Figure 3

The relationship between cell shape and phospho-MLC in individual cells. A) In the absence of any drug treatment, the area of individual cells was linearly correlated with total diphospho-MLC content per cell. The data shown are from a representative experiment indicating measurements on about 150 cells; RFU (relative fluorescent units) B) Inhibiting ROCK by the addition of the indicated concentrations of Y27632 inhibited phospho-MLC in individual cells while preserving the linear relationship between cell spreading and MLC diphosphorylation. Representative data are shown from experiments involving about 150 cells. C) Treatment with Y27632 did not change the distribution of cell areas within the population as indicated by the similarity of the histograms. D) Inhibiting ROCK increased the perimeter of the cells compared to no drug treatment. These data are average values of cellular perimeters determined from three replicate experiments. Statistically significant difference compared to the perimeter of single cells in the absence of inhibitor (p < 0.05) is indicated by '*'. E) Representative cells in the absence of Y27632, F) in the presence of 10 μM Y27632 and G) 100 μM Y27632. Cells were stained with Tx-Red maleimide as described in Methods.