Extended Data Fig. 6: High dimensional analysis of all neurons.

a) Leiden clustering (LC) and UMAP embedding of neurons extracted from the whole time course, labeled by cell type. Circles indicate the 3 main neuronal subtypes by RTK expression: TrkA⁺;Ret⁺, TrkB⁺, and TrkC⁺, respectively. b) LC and UMAP of the TrkA⁺;Ret⁺ neurons, extracted from a). Three clusters that did not exhibit neuronal markers were removed from the dataset before a final round of LC and UMAP (plot on right). c) LC and UMAP of the TrkB⁺ neurons extracted from a). d) LC and UMAP of the TrkC⁺ neurons extracted from a). Putative phagocytic glia expressing TrkC⁺ could not be removed from TrkC⁺ neurons in previous analytic iterations, but this could be done at this resolution resulting in a ‘cleaned’ TrkC⁺ neuronal clustering and UMAP (plot on right). e) TrkA⁺;Ret⁺, TrkB⁺, and TrkC⁺ UMAP plots colored by age. f) Violin plots of all markers for TrkA⁺;Ret⁺, TrkB⁺, and TrkC⁺ neurons. g) Key markers in our panel that allow identification of somatosensory DRG populations⁵. h) UMAP plots colored by expression for all panel markers for TrkA⁺;Ret⁺, TrkB⁺, and TrkC⁺ neurons. i) TrpM8 transcript data from Usoskin et al.² showing that TrpM8-expressing neurons are a subset of peptidergic nociceptors. j) UMAP plot of TH⁺ cells that were extracted from Fig. 4a (Cluster 6) and reclustered. k) UMAP plot of the TH⁺ cells from P4 overlaid on the grayed out UMAP plot from j). These were the cells used in the comparison to the Usoskin et al² transcript data in Fig. 4i-n. l) UMAP plot from j) colored by age. m) Violin plot of marker expression of all clusters from j). n) Comparison of TH⁺ C-LTMR transcripts expression to protein expression for all markers in the mass cytometry panel². In both cases, transcript or protein expression in C-LTMRs was normalized to all nociceptors.