N-cad regulates PHGG cell cohesion during migration. (A) Number of single cells migrating ahead of the migration front after 24 h migration on laminin. (B and C) Images and quantification of cell-cell connections at the migration front. Scale bar, 20 μm. Error bars show mean ± SEM. (D–I) Cell tracking. Dendra2-histone H2B-expressing control or N-cad shRNA cells migrating on neurons or laminin for 16 h. Cell tracks (D and G) and box and whisker plots (E, F, H, and I) indicating median, quartiles, and range. Ordinary one-way ANOVA Holm-Šídák’s multiple comparisons tests. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (A)N = 12 spheroids. (C)N = 16–26 spheroids, three experiments. (E and F)N = 42–102 cells, three spheroids. (H and I)N = 49–70 cells, three spheroids.