Fig. 3

Predicted diseases from the CNS-phosphoproteome of GIFN39 mice and associated clinical and pathology outcomes. A Activation state of diseases and disordered enriched from the regulated phosphoproteome of GIFN39 mice. Positive score (red) indicates increased activity and negative score (blue) indicates decreased activity of the clinical and pathological signs of disease. B Progressive reduction in survival of GIFN39 mice (total n = 317) compared to WT littermates (total n = 376), P < 0.001 by log-rank test. Data presented as mean ± 95% confidence interval (shaded). C Body weight, D length of mice from nose to tail base and E wet brain weight was reduced in male GIFN39 mice compared with WT mice and with age (n = 4–13 per genotype per age). Data presented with mean ± SEM. * P < 0.05, ** P < 0.01 and **** P < 0.0001 between indicated samples as determined by two-way ANOVA with Tukey’s post-test. n.s.: not significant. Neuropathology was investigated in the cerebellum of 8 and > 16-week-old GIFN39 mice and WT littermates. (F) Dual Iba1 (brown/black) and Alizarin Red S (ARS; red; indicated by arrowheads) stain for microglia and calcium deposits. (G) GFAP stain for astrocytes. (H) CD3 stain for T cells, indicated by arrowheads. (I) Neurofilament stain to reveal neurons. (J) Cleaved caspase-3 stain for apoptotic cells indicated by black arrowheads. Red arrowhead indicates an aneurysm. Representative immunohistochemical and histological stains of cerebella of WT and GIFN39 of 8-week-old and 16-week-old mice (n = 4 mice per genotype per age). ML: molecular layer, GCL: granule cell layer and WM: white matter