Extended Data Fig. 1: Transcriptomic analysis of PBN cells.

(a) UMAP projection of all 76,201 PBN cells analyzed by spatial transcriptomics, with each cell colored by its assigned cluster. This clustering reflects only the transcriptional similarities. (b) UMAP embedding by glial identity. Cells expressing glial markers are shown in green. (c) UMAP embedding showing the normalized expression level of Aqp4, an astrocyte marker. Color scale maximum: 16. (d) UMAP embedding showing the normalized expression level of Tmem119, a microglia marker. Color scale maximum: 10. (e) UMAP embedding showing the normalized expression level of Olig2, an oligodendrocyte marker. Color scale maximum: 11. (f) (top) UMAP embedding showing the normalized expression level of Slc17a6, the gene encoding VGLUT2 as a marker of excitatory neurons in all PBN cells. Color scale maximum: 15. (bottom) UMAP embedding in re-clustered neurons showing the normalized expression level of Slc17a6. Color scale maximum: 2.35. (g) (top) UMAP embedding showing the normalized expression level of Slc17a7, the gene encoding VGLUT1 as a marker of excitatory neurons in all PBN cells. Color scale maximum: 51. (bottom) UMAP embedding in re-clustered neurons showing normalized the expression level of Slc17a7. Color scale maximum: 2.35. (h) (top) UMAP embedding showing the normalized expression level of Slc32a1, the gene encoding VGAT as a marker of inhibitory neurons in all PBN cells. Color scale maximum: 69. (bottom) UMAP embedding in re-clustered neurons showing the normalized expression level of Slc32a1. Color scale maximum: 3.15. (i) Dot plot showing a differentially expressed gene from each neuronal cluster. The size of the dot represents the number of cells within that cluster expressing the gene and the color of the dot represents the mean expression of that gene. (j) Dot plot showing the expression of genes previously identified as markers for populations of neurons that are involved in pain behaviors across each neuronal cluster.