Fig. 4 | Nature Communications

Fig. 4

From: Matrix mechanical plasticity regulates cancer cell migration through confining microenvironments

Fig. 4

In highly plastic ECM, cell-generated forces displace the matrix plastically to facilitate invasion and migration. a Probability of cell migration, with the indicated vehicle alone or inhibitor, added to the media. Drug/antibody concentrations used to inhibit/block respective pathways were: 1 μg mL−1 monoclonal β1 integrin-blocking antibody (β1 integrin), 70 μM NSC23766 (Rac1), 10 μM Y-27632 (ROCK), 100 μM CK-666 (Arp 2/3), 50 μM Blebbistatin (myosin II), and 2.5 μM Latrunculin-a (F-actin). Cells from R = 3 biological replicate experiments, bars indicate mean probabilities, and error bars indicate 95% confidence intervals. b, c Images from confocal time-lapse studies of representative RFP-LifeAct MDA-MB-231 cells, encapsulated with fluorescent beads and stimulated with 50 ng mL−1 EGF, depicting b, a cell extending a protrusion, and c, a cell migrating. Bead displacements obtained from a single z-plane and time points shown were used to inform models of the matrix displacement field, which is superimposed over a heat map illustrating displacement magnitudes and directions as calculated. Scale bar is 20 μm. d Average matrix displacement map, which incorporates information from N = 16 cells from R = 3 biological replicate experiments. All centroids were located at the red indicator, and all maps were rotated such that the cells migrated to the right. e, f Maximum bead displacements observed around migrating cells for conditions in which cell migration was observed, and around stationary cells when cell migration was absent. e Maximum bead displacements around cells in HP IPNs for indicated conditions. Statistically significant differences are indicated (**P < 0.01, ****P < 0.0001, ANOVA; ns not significant). f Maximum bead displacements around cells in HP, MP, and LP IPNs (ANOVA). g, h Cells in HP IPNs made with fluorescein-conjugated alginate. g Matrix is densified around protrusions (top yellow arrow), and densification persists after protrusions retract (bottom yellow arrow). h Migrating cells leave lasting channels. The fluorescence intensity signal across the migrating cell’s path (line marked A  to B) was measured. i Three hours after cells were lysed and actin networks were depolymerized, similar channels remained, with their intensity profiles as shown. For gi, scale bar is 10 μm

Back to article page