Fig. 2

In the ventral horn of the spinal cord EfnA5 is mainly expressed in neurons. RNAscope in situ hybridization was performed to determine cell type-specific expression of EfnA5. A total of 15–18 images of different lumbar spinal cord ventral horns from 3 different mice were analysed for every condition. a Representative images show lumbar spinal cord ventral horns of SOD1^WT and SOD1^G93A mice at 135 days of age that were stained with probes against EfnA5, Syp and Slc1a3. Hoechst was used as a counter stain for nuclei. Scale bar = 50 μm. b Percentage of neurons (Syp + cells) that stained positive for EfnA5 were quantified and shown as percentage of the total Syp + cells. c Glial cells (Syp- cells) and d astrocytes (Syp-, Slc1a3+ cells) positive for EfnA5 were quantified and represented as percentage of the total amount of glial cells. Single-cell EfnA5 expression was quantified by measuring EfnA5 puncta density (dots/μm²) in e neurons, f glial cells and g astrocytes. Data represents mean ± SEM, and different conditions were compared with a two-tailed t-test: * P < 0.05; ** P < 0.01 as compared to SOD1^WT. b and e Data represents mean ± SEM, and different conditions were compared with a two-way ANOVA with Tukey’s multiple comparisons test: * P < 0.05 as compared with SOD1^WT; ^## P < 0.01 as compared with 150–400 μm² neurons