Adhesion dynamics is spatiotemporally correlated with an increase in membrane tension in a vinculin-dependent manner. (A) Representative images in late P2 of vinculin KO cells transfected with vinculin head and vinculin full. Bars, 10 µm. (B) Time-lapse images at the transition T of focal adhesions dynamics from A for vinculin head (1) and vinculin full (2). (C) Histogram of vinculin-containing adhesion sizes after 15-min spreading in vinculin full (red) and vinculin head (blue) cells. (D) Histogram of the shift in adhesion sizes from vinculin head to vinculin full. (E) Plot of the tether force values for vinculin full (red box) and vinculin head (blue box). **, P < 0.01 in t test statistics. Blue and red boxes extend from the 25th to 75th percentiles, with a black line at the median and a black cross at the mean; black whiskers extend from 5th to 95th percentiles for both conditions; values outside these ranges are plotted as individual points. The procedures in A–D were repeated for at least 15 different cells in each experimental condition. All cells showed behaviors similar to those represented in the figure. The procedures in E were repeated for at least 60 different cells of each experimental condition.