Fig. 2. Augur identifies neuron subtypes that enable walking after paralysis.

a, Top, single-nucleus RNA-sequencing experimental design to prioritize neuron subtypes recruited by TESS. Middle, chronophotography of mice in the presence or absence of TESS and monoaminergic agonists. Bottom, stick diagram decompositions of right leg movements; leg endpoint trajectory with acceleration at toe-off; activity of extensor and flexor muscles of the ankle. b, Principal component analysis (n = 3 mice) of gait parameters for each condition (small circles). Large circles show the average per group. c, Bar plot shows the average scores on principal component 1 (PC1), which quantify the locomotor performance of paralyzed mice (n = 3) and mice walking with TESS (n = 3). d, Uniform manifold approximation and projection (UMAP) visualization of 18,514 nuclei, revealing the six major cell types of the mouse lumbar spinal cord. e, UMAP visualization of 6,035 neurons subjected to an additional round of sub-clustering and the 39 identified neuron subtypes. f, UMAP visualization of 6,035 neurons, colored by Augur cell type prioritization (AUC). The seven prioritized neuron subtypes with the highest AUCs are highlighted. g, Monosynaptically restricted anterograde tracing in Vsx2-Cre mice reveals V2a interneurons densely innervating motor neurons (ChAT). Similar results were obtained from three independent experiments. h, Dot plot showing expression of the immediate early gene Fos in neuron subtypes prioritized by Augur. i, Confirmation of colocalization of V2a, V1/V2b, and Spp1 marker genes (Vsx2, Slc6a5, and Spp1 respectively) and Fos by RNAscope in situ hybridization. Schematic indicates location of imaging for each marker within the spinal cord to aid specificity. Similar results were obtained from two independent experiments.