Fig. 5

B cells increase in prevalence over time following SCI and contain multiple subtypes. A Immunohistochemical staining of B cells in spinal cord tissue 42 dpi, stained with B220 (Green), Tuj1 (Red), and DAPI (Blue). Large and small dashed boxes indicate the regions shown in high power in A’ and A,” respectively. Scale bars: A 500 μm, A’ and A” 100 μm. B Quantification of number of clusters identified in SCI animals over time following injury. Each dot represents an individual animal. Error bars represent SD. C Quantification of the area of clusters per section, represented in pixels. D Isolated and re-clustered UMAP of clusters 2, 3, and 7 identifies four subclusters of B cells. E UMAP plot highlighting cells isolated from each timepoint, top is 0 dpi, middle is 7 dpi, and bottom is 60 dpi, demonstrating most B cells were isolated at 60 dpi. F Dot plot demonstrating relative abundance and expression of the Ig class detected, of selected markers of early B cells, and selected MHC-I and MHC-II genes. G Schematic showing the developmental lineage of B cells including important events along their development. H Heatmap of re-clustered data, and genes identifying them as a subtype identified by Jensen [57]. I UMAP of B cell clusters, with cells identified by developmental stage