FIGURE 1:

WT vinculin recruitment, but not paxillin recruitment, depends on substrate stiffness and cytoskeletal tension. (A) Vinculin (green) and (B) paxillin (red) on mPADs of 3, 5, and 14 kPa in the presence or absence of 20 µM blebbistatin for 1 h. White arrowheads, indicate FAs. Scale bar, 10 µm. Effects of substrate stiffness (C, D) and blebbistatin treatment (E, F) on vinculin and paxillin FA area plotted as box-whisker plots (median and 10th, 25th, 75th, and 90th percentiles; n > 25). For WT vinculin FA area, p < 0.01 for one-way ANOVA for substrate stiffness. ^#p < 0.01 vs. 3 kPa, ^##p < 0.05 control vs. blebbistatin. (G) Effects of substrate stiffness and actomyosin contractility on vinculin/paxillin intensity ratio at single FAs (n > 25; mean ± SD). *p < 0.01 vs. 3 kPa, **p < 0.01 vs. respective WT control, and ^&&p < 0.01 vs. 5-kPa WT control.