Molecular and temporal profile of macroglial heterogeneity acutely after SCI. (A) UMAP plot of all macroglial cells from uninjured spinal cord and 1, 3, and 7 dpi. Ependymal cells, astrocytes, OPCs and oligodendrocytes (Fig. 1 A) were extracted, reclustered, and reembedded in new UMAP coordinates. Cells are colored by macroglia subtype as shown in legend on right. Cell counts in parentheses. Subtypes were annotated using a combination of DEG, GO enrichment analyses, canonical markers, and transcription factor expression (Figs. S4 and S5). (B) UMAP of macroglial cells split by each time point. Cells are colored by neighborhood density in UMAP space to illustrate shifts over time. Darker red colors indicate greater cell density in the UMAP or gray if not from indicated time point. (C) Proportions of subtypes among astrocytes and ependymal cells and (D) proportions of oligodendrocyte-lineage subtypes at each time point. (E) Dot plot of marker genes that differentiate macroglial subtypes. Color of dots represent z-scored expression level and size of dots represent percent of cells with at least one UMI detected per gene. Inset stars indicate statistically significant greater expression compared with all other cells combined (adjusted P value < 10⁻¹⁰, Wilcox rank sum with Bonferroni correction; Materials and methods). (F) GO biological process terms associated with the top DEGs between time points for astrocytes (left) and OPCs (right). OPC-A and OPC-B were combined for this analysis. (G) Violin plot of expression of common axon growth inhibitors between astroependymal cells, astrocytes, and OPC subtypes. Expression levels are log-normalized counts along right axis. Assemb, assembly; Biogen, biogenesis; Div, dividing; Oligo, oligodendrocyte; OPC, oligodendrocyte progenitor cell; Organiz, organization; Proc, process; Prop., proportion; Uninj, uninjured.