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Fig. 6. Quantification of the effects of mutation at Tyr-397 of FAK/YCam on FA disassembly after nocodazole washout. (A) In FAK/YCam-transfected cells, cellular motility was clearly apparent (directions is indicated by the white arrow), during the 1 hour recovery period after nocodazole (see supplementary material Movie 1), unlike Y397F-FAK/YCam-transfected cells (see supplementary material Movie 2). Scale bar: 10 µm. (B) FA turnover was also inhibited in Y397F-FAK/YCam cells compared with FAK/YCam. Left images at t=0, centre images at t=60 minutes; right images are the result of subtraction of the images at t=60 from those at t=0. In the subtracted images, disassembled FAs are black, newly formed FAs are white and constant FAs are grey. (C) Disassembled, newly formed and constant (immobile) FAs plotted as a percentage of the total FAs in FAK/YCam and Y397F-FAK/YCam cells during recovery from nocodazole treatment. Values are the mean of four separate experiments. (D). Migration speed of FAK/YCam and Y397F-FAK/YCam cells was assessed using a wound-healing model (see Materials and Methods). Y397F-FAK/Ycam cells (n=120) had a significantly decreased migration speed compared with FAK/Ycam cells (n=120).





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