FIGURE 4.

Cell spreading assay. (A) MDCK cells were trypsinized, re-seeded on a fibronectin (0.1 μg/ml, Sigma) coated coverslips were allowed to spread at 37.5°C for required time (1–4 h) in the medium containing either 10 μM of cisplatin, 10 μM of HNK or 10 nM latrunculin B. F-actin was labeled with Phalloidin-Alexa 594. (B) Cell spreading ability was quantitatively accessed by a self-defined “cell-spreading index”. Cell spreading index was then defined as the ratio between total cell surface and cell nucleus surface and around 100 cells/group were measured. Within a 4 h cell spreading time, control cells spread and expanded 2.31-fold when compared with 1 h. When cells were spreading in the presence of latrunculin B (Lat. B), no expansion of cell surface was measured. When cells were allowed to spread in the presence of cisplatin, a 1.5-fold increase in cell surface area was measured; however, this ratio was not significantly differed from negative control (Lat. B) group. A significant increase of cell surface (1.89-fold) was measured the 10 μM of HNK was present in the medium. Images presented were representative images. a,b indicated significant (p < 0.05) difference between groups. NS, non-statistically different.