E-cadherin impairs actomyosin cable formation and promotes pMLC accumulation at E-cadherin–mediated cell–cell contacts during SDF1 chemotaxis. (A) Immunofluorescence staining of mesenchymal and epithelial-like NC clusters in the absence of external chemotactic cues, labeled for pMLC, E-cadherin, and nuclei. Zoom 1 shows the cluster periphery; Zoom 2 shows the central region of the same cluster. (B) Quantification of pMLC fluorescence intensity in the periphery and center of NC clusters shown in A (n = 12 clusters). (C) Quantification of colocalization between pMLC and E-cadherin using Mander’s coefficient in clusters without external cues (n = 10 clusters). (D) Immunofluorescence staining of mesenchymal and epithelial-like NC clusters during chemotaxis toward SDF1, labeled for pMLC, E-cadherin, and nuclei. Zoom 1 shows the cluster periphery; Zoom 2 shows the central region of the same cluster. Scale bar: 50 µm. (E) Quantification of pMLC fluorescence intensity in the periphery and center of clusters shown in D (n = 10 clusters). (F) Quantification of colocalization between pMLC and E-cadherin during SDF1 chemotaxis using Mander’s coefficient (n = 10 clusters). Colocalization is significantly higher in epithelial-like clusters. Each dot represents one cluster. Data are representative of at least three independent experiments. Error bars show mean ± SEM. Each dot represents one cluster. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparisons posttest. ****P ≤ 0.0001; n.s., not significant.