FIGURE 5:

Expression of constitutively active CA-Vav2 promotes PKL localization to adhesions. (A) HT1080 cells transfected with GFP-PKL or GFP-PKL along with HA-CA-Vav2 were spread on FN in the absence of EGF for 30 min and then stained for paxillin. Images are contrast enhanced to equal degrees for presentation. Scale bar, 10 μm. (B) Mean Pearson's r between GFP-PKL and paxillin per cell was calculated using ImageJ. (C) HT1080 cells transfected with either GFP alone or GFP along with HA-CA-Vav2 were spread on FN in the absence of EGF for 30 min and then stained for paxillin and PKL/GIT1. Scale bar, 10 μm. Line profiles through adhesions demonstrate increased intensity of PKL/GIT1 in paxillin-positive adhesions. The average focal adhesion size per cell (D) and the average ratio of PKL/GIT1 intensity to paxillin intensity in adhesions per cell (E) were quantified in background-subtracted raw images using Image J. Values are means ± SEM for three experiments and at least 10 cells per experiment. Significance was determined using Student's t test. *p < 0.05, *** p < 0.005.