Vinculin knockdown induces accumulation of NSCs nuclei at the basal side of the neuroepithelium before affecting the actin cytoskeleton structure. (A) Scheme of the DNA vector and the time schedule used for the experiments shown Fig. 1 B; hpe, hours post-electroporation. (B) Images of transverse sections of chicken NTs transfected at stage HH12 with shControl or shVCL for 24, 36, and 48 h stained with phalloidin (actin, gray scale). GFP indicates transfection (green). Scale bar = 50 μm. (C) Time schedules used for the experiments shown Fig. 1 D. (D) Images of transverse sections of chicken NTs transfected at stages HH18 or HH23 with shControl or shVCL for 24 h stained with phalloidin (actin, gray scale). GFP indicates transfection (green). Scale bar = 50 μm. (E) Drawing illustrating the coordination that exists between INM, cell cycle, and neuron delamination in chicken NT. (F) Dot plot showing the apicobasal position of nuclei from NTs transfected at stage HH12 for 24 h with shControl or shVCL. The black bars indicate the median. t test, n = 17 slices from two independent experiments using at least three embryos per condition. (G) Representative images from those used to generate the plot shown in 1F. GFP indicates transfection (green). Scale bar = 10 μm. (H) Images of NTs transfected at stage HH12 for 24 with shControl or shVCL and stained with antibodies against Sox2 (neural progenitors, green), HuC/D (neurons, magenta). GFP indicates transfection (blue). Sox2 and HuC/D channels are shown separately in green and gray scale, respectively, at the right hand of the panels. Scale bar = 50 μm. (I and J) Quantification of the percentage of Sox2⁺ or HuC/D⁺ cells relative to GFP cells in the NTs shown in Fig. 3 H. Mean ± SEM. t test, n = 14 sections for control and 13 sections for shVCL, from two independent experiments using at least three embryos per condition. ns = non-significant, *** = P < 0.001.