Fig. 2: Increased senescent markers and SASP proteins characterize aged lesion after demyelination.

a Schematic of MERFISH spatial transcriptomics. b Demyelinated lesions were traced in young (3 months) and aged (18-22 months) mice at 5 dpl. c UMAP visualization of lesion cell types in young and aged lesions (n = 2 per group). Uniform manifold approximation and projection (UMAP) visualization of young and aged lesions by cell types. d Comparison of senescent cells in pooled lesions vs naïve non-lesion controls regardless of age (n = 4 per group; each n denotes biologically independent replicate lesion or non-lesion from the same mouse). e Heatmap depicting top differentially expressed genes between young and aged lesions at 5 dpl. Color represents LogFC relative to young lesion and * indicates significantly differentially expressed. f Dot plot describing senescent gene expression across pooled lesion and non-lesion (n = 4 per group). g Dot plot describing senescent gene expression across young and aged lesions (n = 2 per group). h Heatmap depicting percentage of cell types observed to be senescent in young and aged lesion and non-lesion. Color represents % of cells senescent. i Comparison of percentage of senescent microglia and macrophages in pooled lesion and non-lesion regardless of age (n = 4 per group; each n denotes biologically independent replicate lesion or non-lesion from the same mouse). j Heatmap depicting SASP (senescence associated secretory phenotype) protein levels in young and aged lesions relative to young non-lesioned white matter at 5 dpl (n = 3 per group). Color represents LogFC relative to young non-lesion and * indicates significantly differentially expressed. Data are mean ± SEM (d, i). P-values derived from two-sided student’s t-test (d, i), two-way ANOVA with FDR corrected multiple comparisons (j) or the two-sided Wald test (e). Select figures created in BioRender (2025) (a) https://BioRender.com/j71l219. Source data are provided as a Source Data file.