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. 2018 Apr 10;114(7):1680–1694. doi: 10.1016/j.bpj.2018.02.019

Figure 6.

Figure 6

FRET-FRAP assay shows that force-sensitive vinculin dynamics depend on talin and actin interactions. (A) Correlation between recovery half-time and FRET efficiency for WT vinculin corresponds to the force-stabilized state (n = 32, p < 0.005). (B) Correlation between recovery half-time and FRET efficiency for VinTS A50I corresponds to the force-destabilized state (n = 21, p < 0.05). (C) No detectable correlation between recovery half-time and FRET efficiency was observed for VinTS I997A (n = 18, p = 0.88). (D) Y-27632 treatment reverses the relationship between recovery half-time and load for WT vinculin, corresponding to the force-destabilized state (n = 24, p < 0.05). (E) Y-27632 treatment does not affect the relationship between recovery and load for VinTS A50I, which remains in the force-destabilized state (n = 16, p < 0.01). (F) Y-27632 treatment does not affect the relationship between recovery and load for VinTS I997A, with turnover still insensitive to load (n = 16, p = 0.98). The p values indicate the results of a t-test comparing the regression slope to zero. (Vertical dotted lines) Previously established zero-load is shown.