Fig. 1

Canonical senescent markers increase in CNS meningeal immune cells following EAE induction. A Schematic of EAE experimental timeline. Spinal cord tissue from mice with a score of 3 was collected at 28 days post-immunization (dpi). B Senescence-associated β-gal staining of naïve and EAE lumbar spinal cord shown with black arrows. C Naïve and EAE spinal cords stained with p16 and p21 are shown with white arrows, Iba1, and Hoechst nuclear stain. D Immunofluorescence (IF) staining of EAE spinal cord grey matter (GM), normal appearing white matter (NAWM), lesion, and leptomeninges with p21, Iba1 marker for microglia or PDGFRβ marker for mural cells, and Hoechst. E Quantification of d (one-way ANOVA with Tukey’s multiple comparison test; F_(4,10) = 45.51, df = 4, p < 0.0001). F IF staining of EAE spinal cord for Ki67 marker for proliferation, and G cleaves caspase 3 (CC3) marker for apoptosis. H IF staining of lumbar EAE spinal cord with p21 and p16 with markers for T cells (CD3), B cells (B220), myeloid cells (CD11b). Data are presented as ± SEM; n = 3 biological replicates, ns = not significant, ****P < 0.0001. Scale bars: b, 100 μm; c, 200 μm; d, f, g, h 20 μm