E-cadherin levels in St13, St17, and St17 + E-Cad NC clusters, and stability of junctional E-cadherin during the chemotactic window in St13 explants. (A) Immunofluorescence images of NC clusters from St13, St17, and St17 + E-Cad NC cells showing junctional E-cadherin distribution. Heatmaps show relative fluorescence intensity, and insets present enlarged views of selected junctions. Scale bar: 100 µm. (B) Fluorescence intensity profiles measured along cell–cell contacts for the conditions shown in A. (C) Quantification of peak junctional fluorescence intensity (n = 10 clusters per condition). (D) Immunofluorescence images of St13 NC clusters fixed at 0, 4, and 10 h after plating, illustrating E-cadherin localization over time. Heatmaps show relative intensity, with insets highlighting junctional regions. Scale bar: 100 µm. (E) Fluorescence intensity profiles along junctions at 0, 4, and 10 h. (F) Quantification of peak junctional fluorescence intensity shows stable E-cadherin levels throughout the 10-h interval (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.