Extended Data Fig. 2: Analysis of PAA gel stiffness and collagen concentration.
From: Self-generated gradients steer collective migration on viscoelastic collagen networks
a. Representative micrographs from PAA gel coated with different concentrations of fluorescently-labeled monomeric collagen or 2mg/ml collagen networks. Scale bar: 30μm. b. Boxplot of mean collagen intensity for conditions shown in a. Each point represents one imaging field, relative to background fluorescence in areas with no coating. For a,b, images/data from n=5 positions per condition from one representative experiment, N=3 independent experiments. One-way ANOVA p=2.10e-11. *p < 0.05, **p < 0.01, ***p < 0.001 for Tukey HSD post-hoc test (p=0.00199, < 0.001, 0.0101, 0.00162, < 0.001, 0.0305, 0.679, 0.00717, < 0.001, 0.121, 0.00186, < 0.001, 0.00674, < 0.001, 0.248). c. Example micrographs of cell clusters plated on PAA gels with different concentrations of monomeric collagen. Scale bar: 50μm. d. Boxplots of mean instantaneous speed and coefficient of persistence for conditions in c. Data represents n=68, 72, 97, 56, 52, 46 clusters, N=3, 3, 3, 3, 2, 2 independent experiments. Data for collagen network and 100μg/ml monomeric collagen is the same data as presented in Fig. 1f. One-way ANOVA p=1.68e-20, 2.42e-25. *p < 0.05, **p < 0.01 for Tukey HSD post-hoc test (p=0.0304, 0.0368, 0.00153, < 0.001, 0.00795, 0.659, 0.0547, < 0.001, < 0.001, 0.0145, < 0.001, < 0.001, < 0.001, < 0.001, < 0.001; p=0.855, 0.773, 0.0490, < 0.001, < 0.001, < 0.900, 0.0518, < 0.001, < 0.001, 0.0383, < 0.001, < 0.001, 0.0253, < 0.001, < 0.001). e. Example micrographs of cell clusters plated on PAA gels of different stiffness (100μg/ml monomeric collagen). Scale bar: 50μm. f. Boxplots of mean instantaneous speed and coefficient of persistence for conditions in e. Data represents n=72, 62, 57, 69, 46, 46, 114 clusters, N=3, 3, 3, 3, 3, 2, 6 independent experiments. Data for collagen network and 100μg/ml monomeric collagen is the same data as presented in Fig. 1f. One-way ANOVA p=6.26e-13, 6.25e-30. *p < 0.05, **p < 0.01, ***p < 0.001 for Tukey HSD post-hoc test (p < 0.001, 0.00688, 0.238, 0.00704, < 0.001, < 0.001, < 0.001, < 0.001, 0.0122, 0.889, 0.564, 0.137, 0.504, < 0.001, < 0.001, 0.0758, < 0.001, < 0.001, 0.0143, 0.00996, 0.700; p= < 0.001, < 0.001, 0.728, 0.850, 0.233, < 0.001, < 0.001, 0.00300, < 0.001, 0.0327, < 0.001, < 0.001, 0.589,0.427, < 0.001, < 0.001, 0.158, < 0.001, < 0.001, < 0.001, < 0.001, 0.0763). g. Representative micrographs of fluorescently-labeled collagen networks with different collagen concentration or polymerization temperature. Scale bar: 100μm. h. Boxplot of mean collagen intensity for conditions shown in g. For g,h, representative images/data from n=4, 3, 3, 3, 3 positions, N=1 experiment. One-way ANOVA p=7.74e-6. i. Boxplots of mean instantaneous speed and coefficient of persistence for cluster migration on collagen network conditions in g. Data represents n=37, 114, 47, 85, 43 clusters, N=2, 3, 6, 3, 2 independent experiments. Data for 2mg/ml 37∘ C is the same data as presented in Fig. 1f. One-way ANOVA p=6.56e-12, 0.000609. *p < 0.05, **p < 0.01, ***p < 0.001 for Tukey HSD post-hoc test (p < 0.001, 0.00918, 0.175, 0.121, < 0.001, < 0.001, < 0.001,0.111,0.206,0.793; p=0.0134, 0.58, 0.812, 0.893, 0.0217, 0.00226, 0.00612, 0.775, 0.468, 0.705).
