Reduction of cytoskeletal tension via ROCK inhibition affects vinculin dynamics only in the presence of vinculin-talin interaction. (A) Representative FAs of cells expressing the VinTS and its mutant variants (VinTS A50I, VinTS I997A) treated with 25 μM Y-27632 are displayed in the acceptor channel during FRAP imaging. Scale bars, 3 μm. (B) Vinculin FRAP recovery curves are shown for cells expressing VinTS, VinTS A50I, or VinTS I997A (n = 24, 16, and 16 FAs, respectively, from four independent experiments). (C) Box-whisker plots are given of the half-time of recovery for the FAs analyzed in (B). No significant difference was detected between groups. (D) Box-whisker plots of the mobile fraction are given for the FAs analyzed in (B). Differences between groups were detected using the Steel-Dwass test (∗∗∗p < 0.001); p values for all comparisons can be found in Table S2.