FAK activity and C-terminal PRR regions are required for FA turnover.

<p>(A) Scratch wound assay performed with FAK−/− fibroblasts expressing GFP or the indicated GFP-FAK constructs. Closure percentage was calculated by the change in area between 2 and 10 h. Experimental values are presented as ± SEM from 3 independent experiments (***p<0.001). (B) Representative image montage (10 to 30 min) from live-cell spinning disc confocal microscopy of FA-localized GFP-FAK after growth media supplemented with 50 ng/ml EGF stimulation. A merged image from the 10/20/30 min time points was pseudo-colored red (10 min), green (20 min) and blue (30 min) respectively to illustrate GFP-FAK localization over time. White regions indicate GFP-FAK localization overlap at 10 to 30 min. Scale is 10 µm. (C) Adhesion lifetime was determined by counting the number of sequential frames (2 min intervals within a 60 min time-lapse) of GFP-FAK FA-associated fluorescence above background. Data is the mean lifetime ± SEM of 150–200 FAs from at least 5 different cells from each of the indicated GFP-FAK reconstituted FAK−/− MEFs (**p<0.01, compared to FAK-WT).