Extended Data Fig. 5: Analysis of focal adhesions and integrins during migration.
From: Self-generated gradients steer collective migration on viscoelastic collagen networks
a. Fixed cell clusters plated on glass or on a collagen network and immunostained for Paxillin with DAPI (DNA) and Phalloidin (F-Actin) counterstaining. Representative examples, N=2 independent experiments. Scale bars: 50μm. b. Single time points from live imaging of A431 cells stably expressing Paxillin-GFP or Zyxin-mCherry, plated on glass or collagen networks. Representative examples, N=3 independent experiments. Scale bar: 50μm. c. Boxplots of mean instantaneous speed and coefficient of persistence for cluster migration on collagen networks with the integrin binding inhibitors Cilengitide and AIIB2 and MMP inhibitors GM6001 and BB94. Data represents n=146, 57, 72, 31, 26, 17, 14 clusters, N=8, 3, 4, 2, 2, 1, 1 independent experiments. Control data is pooled from data presented in Fig. 1f. One-way ANOVAs: p=4.36e-6, 1.76e-18. **p < 0.01, ***p < 0.001 for Tukey HSD post-hoc test (p=0.268, 0.126, < 0.001, 0.00492, 0.390, > 0.9, 0.832, < 0.001, < 0.001, 0.865, 0.447, < 0.001, < 0.001, > 0.9, 0.256, 0.537, < 0.001, < 0.001, < 0.001, < 0.001, 0.243; p=0.0788, 0.0195, < 0.001, < 0.001, 0.88, 0.25, 0.689, < 0.001, < 0.001, 0.130, 0.016, < 0.001, < 0.001, 0.061, 0.00522, 0.268, < 0.001, 0.0148, < 0.001, 0.00246, 0.354). d. Example micrograph from a cell cluster on a collagen network following treatment with the Integrin-β1 functional antibody AIIB2 (10μg/ml). Scale bar: 100μm. HH:MM. See also Supplementary Video 4. Representative image series from n=26 clusters, N=2 independent experiments.
