Extended Data Fig. 2: Generation and analysis of Mpz-enriched single nuclei atlas.

(a) Gating strategy for sorting GFP+ nuclei from Sun1-GFP/+:Mpz-Cre/+ mice. Nuclei were isolated from single cell suspensions containing pooled sciatic nerves from at least 5 Sun1-GFP/+:Act-Cre/+ mice. Purified nuclei were selected based on GFP and Hoechst expression. (b) tSNE plot with 3,047 nuclei from Sun1-GFP/+;Mpz-Cre/+ defines three GFP+ cell types – endoneurial fibroblasts, nmSCs and mSCs. (c) Differential expression analysis from Mpz-Sun1 atlas shows signature gene expression that defines endoneurial fibroblasts, including Col15a1, Lama2 and Egfr. Each column represents a gene and each row represents a nucleus, genes with high expression level are shown in yellow and low expression in purple. (d) GO analysis showing that endoneurial fibroblast markers (122 genes) are enriched for components related to collagen-containing extracellular matrix, extracellular matrix and formation of axon, neurons and synapse, percentage of GO recovery is plotted against the false discovery rate (FDR) and selected GO terms are highlighted in color. (e,f) RNA-FISH analysis of Prx, Ngfr, and Cd34 expression. (scale bar=50um) e, Representative expression of Prx and Ngfr, indicating that myelinating Schwann cells do not express Ngfr. f, Representative expression of Prx and Cd34, indicating that myelinating Schwann cells do not express Cd34. (g) RNA-FISH analysis of Csmd1 and Scn7a. Representative overlapping expression of Csmd1 (green) and Scn7a (red) in adult sciatic nerves (scale bar=50um).