Fig. 6

NAD⁺ supplementation depends on glycolysis to rescue APP transport in NMNAT2 KO neurons. A Representative kymographs from DIV8 ctrl or KO neurons supplemented with Veh (vehicle) or 1 mM NAD⁺ since DIV5. Scale bar, 20 µm. B Percentages of APP-EGFP vesicles moving anterogradely, retrogradely, or pausing in distal axons of DIV8 ctrl and KO neurons supplemented with Veh or NAD⁺. Neurons imaged and statistics: 56 ctrl + Veh, 62 KO + Veh, 17 ctrl + NAD⁺, and 49 KO + NAD⁺, *p = 0.0232, ^##p = 0.0018. Significant comparison to ctrl + Veh and KO + Veh groups was labeled with* and ^#, respectively. C Velocity of antero- and retrograde transport of APP-EGFP in distal axons of DIV8 ctrl and KO neurons supplemented with Veh or NAD⁺. Numbers (neurons imaged) and statistics: 56 ctrl + Veh, 51–58 KO + Veh, 17 ctrl + NAD⁺, and 47–48 KO + NAD⁺ for anterograde velocity, (anterograde) **p = 0.0041, (retrograde) **p = 0.0013, ***p = 0.0002. E Schematic representation of how glycolysis and mitochondrial respiration are affected by Oligo or 2DG(+ Methyl-Pyr) treatment. F Percentages of APP-EGFP vesicles moving anterogradely, retrogradely, or pausing in distal axons of DIV8 ctrl and KO + NAD⁺ neurons in the presence of vehicle, Oligo, or 2DG. Numbers (neurons imaged) and statistics: 56 ctrl + Veh, 39 ctrl + oligo, and 20 ctrl + 2DG, **p = 0.0059; 49 KO + NAD⁺ + Veh, 22–25 KO + NAD⁺ + Oligo,and 15–18 KO + NAD⁺ + 2DG, *p = 0.0291, ^#p = 0.0252. Significant comparisons to Veh-treated and Oligo-treated groups were labeled with * and ^#, respectively. G Velocity of antero- and retrograde transport of APP-EGFP in distal axons of DIV8 ctrl and KO + NAD⁺ neurons in the presence of vehicle (black), Oligo (blue), or 2DG (green). Neurons imaged and statistics: 56 ctrl + Veh, 38–39 ctrl + oligo, 10 ctrl + 2DG, 47–48 KO + NAD⁺, 24 KO + NAD⁺ + Oligo, and 12–13 KO + NAD⁺ + 2DG, **p = 0.0011. All above data represent mean ± SEM, two-way ANOVA with Tukey's multiple comparisons test or Kruskal–Wallis test with Dunn's multiple comparisons test were used, see details in supplemented excel, ****p < 0.0001, ^####p < 0.0001