Fig. 2: Glucose utilization changes from delamination to migration.

a Schematic representation of the metabolic pathways downstream of glucose and glutamine uptakes and their interconnections with the PP and OXPHOS pathways. The main enzymes within each pathway analyzed in this study, as well as the specific metabolic inhibitors used, are indicated. b Scatter dot plot of normalized mRNAs levels of genes encoding glycolytic enzymes in cultured NT explants with quiescent (yellow) and OXPHOS (blue) profiles (left) and in NTs collected from the trunk region of HH12/13 (magenta) and HH14 (green) embryos (right). c–e Effect of inhibitors of glycolysis (2-DG, triangles), OXPHOS (Oligomycin, squares), and PP pathways (6-AN, diamonds) compared with medium without drug (circles) on the metabolic activity of NT explants at the delamination (quiescent, top panels) and migration (OXPHOS, bottom panels) stages: c energetic maps; d scatter dot plots of the OCR/ECAR ratio; e ATP levels. OCR and ECAR were recorded in individual explants before and after inhibitor treatment, and ATP levels were measured after two additional hours in the presence of the inhibitor. f Changes in OCR and ECAR over time of NT explants with quiescent (left) and OXPHOS (right) profiles before and after the addition of 6-AN. Data were collected from at least three independent experiments. g Scatter dot plot of normalized mRNA levels of LDH, PFK, and PRPS in the trunk of embryos 5 h after 6-AN or vehicle (no-drug) injection. Embryos were injected either at HH12/13 (magenta) or at HH14 (green). In b left (n) indicates the number of experiments with the measurements done in triplicates for each gene; in b right and g (n) indicate the number of embryos, with the measurements done in triplicates for each gene. In c–f (n) indicate the number of explants analyzed. Data in b, g were analyzed using an unpaired two-tailed t-test. Data in d, e were analyzed using one-way ANOVA followed by Dunnett’s multiple comparison tests relative to the no-drug condition. Data were expressed as mean values ± s.e.m. in b–e, g and as mean ± s.d. in (f). ns not significantly different, P > 0.05.