Fig. 5

Slingshot migration (SSM) requires matrix adhesion and actomyosin contractility. a, b Migration speed, SSM population (% of cell population undergoing one or more SSM event), and SSM duration (% of tracked time of entire population spent in some phase of SSM) in aligned, intermediate stiffness (6.2 kPa) matrices with a range of cRGD functionalization. n = number of cells per group as indicated within each bar (a); n = 3 fields of view with each field of view containing 10 cells (b). c, e, g Migration speed in aligned, intermediate (6.2 kPa) and high (22.5 kPa) stiffness matrices treated with blebbistatin, Y27632, or calyculin A (n = number of cells per group as indicated within each bar). d, f, h SSM population and slingshot duration in aligned, intermediate stiffness (6.2 kPa) matrices treated with blebbistatin, Y27632, or calyculin A (n = 3 fields of view with each field of view containing 10 cells). Migration speed, SSM population, and SSM duration were quantified over a 6 h time course for cRGD, blebbistatin, and Y27 studies, and quantified over a 1 h time course for calyculin A studies (indicated with ^†). i Representative traction stress maps of NIH3T3s on 7.9 kPa polyacrylamide hydrogels subjected to equivalent perturbations as in a–h (scale bar: 50 μm). j Slingshot population (normalized to control) as a function of peak traction stress determined by traction force microscopy. Dashed line indicates linear correlation with indicated R² and p-value. n = number of cells per group as indicated beside each data point. All data presented as mean ± s.d.; * indicates a statistically significant comparison with p < 0.05 (a–c, e, g: one-way analysis of variance; d, f, h: two-tailed Student’s t-test; j: linear regression)