Figure 3

Histopathological changes in GR50 expressing mice. (A) Kaplan–Meier analysis of survival of GR50-GFP and control-GFP male and female mice over 12 months; n = 37 control-GFP mice, 29 GR50-GFP mice. (B) Representative image of ChAT-staining in the ventral horn of the lumbar spinal cord in female mice at 3 and 12 months. Scale bar = 100 µm. Quantification of chat-positive cells at 3-, 6-, 9- and 12-months. n = 3 mice per time-point, min. 6 fields per mouse for quantification, mean ± s.e.m. Two-way ANOVA, Sidak’s multiple comparisons test. 3-months p = 0.7329, 6-months p = 0.0163, 9-months p = 0.0051, 12-months p = 0.0269. (C) Same as in (B) but for male mice. 3-months p = 0.9997, 6-months p = 0.8340, 9-months p = 0.1198, 12-months p = 0.7119. (D) Representative image of GFAP fluorescence within the lumbar ventral grey matter in female control-GFP and GR50-GFP mice at 3-months-old. Scale bar = 50 µm (E) Quantification of lumbar ventral gray (left) and cortical GFAP expression (right) in female mice at 3-, 6-, 9- and 12-months-old. n = 3 mice per time-point, min 6 fields per mouse for quantification, mean ± s.e.m. Two-way ANOVA, Sidak’s multiple comparisons test. Female Lumbar: 3-months p < 0.0001, 6-months p = 0.2401, 9-months p < 0.0001, 12-months p = 0.8294. Female Cortex: 3-months p = 0.1654, 6-months p > 0.9999, 9-months p = 0.0059, 12-months p < 0.0001. (F) Same as in (E) but for male mice. Male Lumbar: 3-months p = 0.8662, 6-months p = 0.9017, 9-months p = 0.9442, 12-months p = 0.0551. Male Cortex: 3-months p = 0.9886, 6-months p = 0.0559, 9-months p = 0.4431, 12-months p = 0.9192. (G) Quantification of cleaved caspase-3 puncta accumulation within cleaved caspase-3 positive cells of the lumbar spinal cord in female control-GFP and GR50-GFP mice at 3- and 12-months-old. n = 3 mice per time-point, min 6 fields per mouse for quantification, mean ± s.e.m. Two-way ANOVA, Sidak’s multiple comparisons test. 3-months p = 0.9974, 12-months p < 0.0001. Right: representative images of cleaved caspase-3 puncta in control-GFP and GR50-GFP female lumbar spinal cord at 3- and 12-months-old. Scale bar = 20 µm. (H) Same as in (G) but for males. 3-months p = 0.9102, 12-months p = 0.0110. Right: representative images of cleaved caspase-3 puncta in control-GFP and GR50-GFP male lumbar spinal cord at 3- and 12-months-old. Scale bar = 20 µm.