Phosphorylation of S200 and S201 is necessary for mitochondrial arrest in response to energetic stress. (A) Kymographs of hippocampal neurons expressing MYC-hTRAK1 or MYC-hTRAK1^{S200A, S201A}, as well as mito-dsRED and meGFP. Cells were incubated for 2 h with 4 nM AntA or vehicle, and imaging was done on DIV9–11. Mitochondria are magenta, and axons are green. (B) Quantification of kymographs as in A. n = 6–9 axons per treatment from five independent animals. (C) Quantification of kymographs of mitochondrial motility in response to 1 h of 2-deoxyglucose treatment in hippocampal neurons expressing either MYC-hTRAK1 ^{S200A, S201A} or MYC-hTRAK1. 2-deoxyglucose treatment was done in neuron media lacking glucose and mitochondria, and neurites were visualized as in A. n = 6–9 axons per treatment from four independent animals. For all images, scale bars = 20 µm. For all graphs, bars on boxplots show the 10–90th percentile, and error bars on bar graphs show the SEM. P values for B and C were calculated by performing a blocked one-way ANOVA with Tukey’s multiple comparison correction. Select comparisons are shown.