Fig. 2. T474 phosphorylation promotes FERM-mediated autoinhibition of FAK. FLAG-tagged FAK proteins (FLAG-FAK) harboring deletion of the FERM-domain (ΔFERM) or the mutations, Y180A/M183A (YA/MA), V196D/L197D (VD/LD), or F596D (FD) were expressed in HEK293T cells, and the levels of pY397 were assessed by Western blot analysis using pY397-FAK-specific antibodies. The total levels of overexpressed FLAG-FAK were determined using FLAG antibodies (left panel). The band intensities of pY397-FAK were quantified, normalized to that of FLAG, and plotted (right panel). Values shown represent the means+SEM of three separate experiments. *p<0.05; **p<0.01.
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