Fig. 4: Vascular and astrocyte cell subclustering identified subsets of senescent-like cells.

a Vascular cell subset based on snRNAseq initial clustering. b UMAP of vascular subclusters and annotation based on Yang et al.⁶² Heatmap displaying gene module scores between subclusters and Yang et al.⁶². c Percentage of the vascular cell populations by location and pathological stage. d Quantification (%) of SenMayo-defined senescent-like cells in each vascular subclustered population demonstrating the presence of subsets of senescent cells in each subcluster. e Representative multiplex immunostaining showing the co-existence of p16^INK4a+ and p16^INK4a− CD31⁺ vascular cells at the chronic active edge. f UMAP of astrocyte subtypes and annotation by cortex and white matter based on Absinta et al.⁸ and Lerma-Martin et al.¹⁸. g Dot plot of averaged z-transformed gene expression of marker genes for each astrocyte subtype. h Percentage of the astrocyte subclustered populations by location and pathological stage. i Quantification (%) of SenMayo-defined senescent cells in each astrocyte subclusters. j Representative multiplex immunostaining showing the co-existence of p16^INK4a+ and p16^INK4a− Vimentin⁺ inflammatory astrocytes at the chronic active lesion edge. Cl cluster, WM white matter, CX cortex, AIMS astrocytes inflamed in MS.