A-Wound healing assays in A549 cells stably expressing wild type CRK-II (WT) or CRK-II phosphomimetic (Ser41Asp) mutants following siRNA mediated PAK1 silencing.

<p>Endogenous CRK is silenced in all conditions by siRNA. B- Measurement of wound surface area 24 hours after the establishment of the wound among the above mentioned groups. The average is calculated following measurement of three separate experiments. (2 tailed student’s t-test: ** P<0.01; *** P<0.001; error bars represent ± standard deviation). C- Invasion assays following incubation of stably transfected A549 cells expressing wild type CRK-II (WT) or CRK-II (Ser41Asp) mutant. Each cell line was treated with PAK1 siRNA or scrambled sequence siRNA. 1×10⁵ cells were incubated over night as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042012#s4" target="_blank">Methods</a> section and stained at 36 hours after incubation. D- Quantitative measurement of the invaded cells. Five separate sections of the invaded cells in each Matrigel membrane were counted. (2 tailed student’s t-test: ** P<0.01; *** P<0.001; error bars represent ± standard deviation).