Fig. 8: β-catenin and glutaminase co-distribute in nuclei of delaminating NCCs.

a Confocal views of β-catenin and GLS distribution in delaminating and migrating NCCs after 5 h in culture in control medium with glucose and glutamine and in medium containing 50 µM CB-839. Images show GLS and β-catenin immunostainings (green and magenta, respectively), Dapi visualization of nuclei (blue), and DIC images of the cell structure (gray). Arrowheads point at β-catenin and GLS accumulation in nuclei of delaminating NCCs and arrows point at β-catenin accumulation in cell-cell contacts in NCCs. Bar = 20 µm. b, c Scatter dot plots of the percentage of NCCs with nuclear β-catenin (b) and with nuclear co-localization of β-catenin and GLS (c) among delaminating (green symbols) and migrating (magenta symbols) NCCs treated or not with CB-839 at 50 µM. d Truncated violin plots of the total fluorescence intensity of β-catenin immunostaining related to the nucleus area in delaminating (green symbols) and migrating (magenta symbols) NCCs with or without CB-839 at 50 µM. e Confocal images of cross-sections through the NT at the level of NCC delamination immunolabeled for β-catenin (red) and GLS (green) and stained with Dapi (blue): top and bottom panels show embryos treated for 5 h with control siRNAs (n = 3) and with siRNAs against GLS (n = 3), respectively. Insets show details of delaminating NCCs in the dorsal NT. Arrowheads point at the nuclei of delaminating NCCs. The area of premigratory NCCs in the dorsal NT is delineated with a white line. Bar = 50 and 10 µm in insets. Images in a, e are from representative experiments. In b, c, (n) indicate the number of explants analyzed, and in d (n) indicate the number of nuclei analyzed. In b–d, data were analyzed using one-way ANOVA followed by Dunnett’s multiple comparison test relative to the no-drug condition. Data were expressed as mean values ± s.e.m. in (b, c) and as mean and first and third quartiles in (d). ns not significantly different, P > 0.05. ec ectoderm.