Progressive changes in cell dispersion and E-cadherin levels across developmental stages of NC explants. (A) Representative frames from time-lapse recordings of NC clusters dissected from St13, Stage 15, and St17. Clusters were plated on fibronectin and imaged over 6 h. Shown are the initial (0 h) and final (6 h) frames. Scale bar: 50 µm. (B) Visualization of dispersion at 6 h based on Delaunay triangulation of nearest-neighbor nuclei. Triangles are color-coded according to area. (C) Quantification of total dispersion area per cluster (n = 10 clusters per condition). (D) Representative immunofluorescence images showing E-cadherin localization in St13, Stage 15, and St17 NC clusters. Heatmaps indicate relative fluorescence intensity for visualization. Insets show magnified views of junctional regions extracted from the heatmap images. Scale bar: 100 µm. (E) Fluorescence intensity profiles along cell–cell junctions for NC clusters at St13, Stage 15, and St17. (F) Quantification of maximum junctional fluorescence intensity values (n = 10 clusters per condition). Each dot represents one cluster. Data are from at least three independent experiments. Error bars indicate mean ± SEM. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparisons posttest. ****P ≤ 0.0001.