FIG. 3.

(a) Representative bright-field images of highly metastatic PC3, moderately metastatic DU145, and normal RWPE-1 prostate cells in control iso-osmotic (ctrl) and hypo-osmotic (hypo) media for 12 h. Arrows indicate blebs, scale bars represent $40\mu \mathrm{m}$. Images were linearly contrast enhanced by stretching gray scale values from the ranges [11, 199] (ctrl) and [0, 153] (hypo) to [0, 255] for PC3; [0, 130] (ctrl) and [22,210] (hypo) to [0,255] for DU145; and [0,163] (ctrl and hypo) to [0,255] for RWPE-1. Displayed images are cropped. (b) The average percentage of cells $\pm$ SD that formed blebs in hypo-osmotic media for 12 h in comparison with control, $\mathrm{\#}\mathrm{p}=0.036$, $\mathrm{\#}\mathrm{\#}\mathrm{p}=0.02$, *$\mathrm{p}=0.0075$, **$\mathrm{p}=0.0069$, ***$\mathrm{p}=0.0052$, one-way ANOVA followed by Tukey’s post hoc test. Highly metastatic PC3 control $\mathrm{n}=582$, PC3 hypo-osmotic $\mathrm{n}=527$, moderately metastatic DU145 control $\mathrm{n}=1169$, DU145 hypo-osmotic $\mathrm{n}=255$, normal RWPE-1 control $\mathrm{n}=507$, RWPE-1 hypo-osmotic $\mathrm{n}=494$. (c) Average viability $\pm$ SD of hypo-osmotically treated (12 h) and control cells determined by Trypan blue stain exclusion. N.s. indicates no statistical significance, two-sample Kolmogorov–Smirnov test. Averages were obtained from 3 independent experiments.