Fig. 3: snRNA-seq analyses of OL upon rhEPO/placebo treatment.

a Two-dimensional UMAP showing the 5 clusters obtained using unbiased transcriptome analysis of 12043 single nuclei, derived from hippocampi of mice treated with rhEPO/placebo. Each dot represents a single nucleus. Clusters were manually annotated by cross-referencing established markers (subfigure b). MOL1/2 were separated based on their calculated pseudotime (MOL2 pseudotime>MOL1 pseudotime, Wilcoxon rank sum test, p < 0.001). b DotPlot illustrating the expression pattern of established OL markers. Average expression denotes the RNA expression of each marker normalized within respective clusters. These markers are used to confirm the identity of the clusters. MOL1 and MOL2 exhibit a higher level of Apc expression separating them from NFOL lineage. c Heatmap representing the differentially expressed genes (DEG, N = 637) upon rhEPO/placebo for each cluster. The table beneath the plot shows the numbers of DEG for each cluster. DEG are represented by their average log2 fold change (minimum average log2FC ± 0.25; adjusted p value ≤ 0.05, ‘bimod’ test, multiple correction done using Bonferroni). Red represents a higher expression value on rhEPO treated samples when compared to placebo samples, while blue represents a lower expression value for the same comparison. The full list of DEG is provided in Supplementary Table 1. d Multiple violin plots showing the expression level and distribution of DEG from the OPC cluster, which belong to the Gene Ontology (GO) term gamma-aminobutyric acid signaling pathway (GO:0007214). Note that all DEG from this GO term were downregulated under rhEPO. P-values calculated using the ‘bimod’ test and adjusted using Bonferroni. Gray represents the expression values from placebo samples and red the ones from samples treated with rhEPO. e Multiple violin plots showing the expression level and distribution of DEG from the OPC cluster, which belong to the GO term glutamate receptor signaling pathway (GO:0007215). Again, all DEG from this GO term were downregulated under rhEPO. P-values calculated using the ‘bimod’ test and adjusted using Bonferroni. Gray represents the expression values from Placebo samples and red the ones from samples treated with rhEPO. f Selected GO terms enriched significantly (over-representation analysis, adjusted p value < 0.05, Benjamini-Hochberg procedure, calculated using clusterProfile⁵⁷) by the analysis of those genes that constructed the rhEPO exclusive regulons (see SCENIC analysis in Methods section). The full list of regulons and GO terms is provided in Supplementary Tables 3 and 4. Source data are provided as a Source Data file.