Fig. 7

Overexpression of ILK L207W causes defects in cell spreading and migration. a HEL cells expressing EGFP-ILK WT or EGFP-ILK L207W were stimulated with 800 nM PMA for 10 min, spread on 20 μg ml⁻¹ fibrinogen for 1 h, fixed and stained with Alexa 647-phalloidin to visualize actin and anti-vinculin antibodies to mark focal adhesions. Transfected cells were identified with EGFP fluorescence. (Top panels) WT EGFP-ILK promotes formation of F-actin stress fibers. By contrast, the stress fibers were substantially disrupted in cells expressing the mutant ILK (bottom panels). Scale bar, 10 µm. b Quantitative analysis of inhibition of cell spreading by ILK L207W as compared to the WT ILK. The areas of EGFP-positive cells were measured using ImageJ software. *P ≤ 0.001. In total, 150 cells were quantified in each sample. The box of the boxplot illustrated the upper and lower quartile for each population (EGFP vector, EGFP-ILK-WT and EGFP-ILK L207W). Median of spreading area is marked by a horizontal line within the box. The attached whisker indicates the range, and the discrete points (•) are the outliers.  c Overexpression of WT ILK but not the L207W mutant in HeLa cells promotes cell migration significantly. Images were captured after 10 h migration, under 10x magnification. d Quantitative change of cell migration with WT ILK versus ILK L207W mutant. The relative fold change of migrated cells was calculated from average of five randomly picked fields for each insert. The results were obtained from three independent experiments. Values are given as mean ± SD