Figure S1. Measurements of the internal diameters of axons. The shortest side of a box (dashed line) surrounding an axon was measured as the internal diameter of the axon (arrow). Scale bars, 1 μm.

Figure S2. Low-power field view of double immunofluorescence staining of the lumbar cord with anti-neurofilament and anti-rhGAL-1 antibodies. In SOD1G93A/Lgals1+/+ mice, large aggregates with neurofilament IR (green, arrows) were often observed especially at the anterior root exit zone, and which were mostly galectin-1 IR positive (red). In SOD1G93A/Lgals1−/− mice, aggregates with neurofilament IR were also found (dashed arrows) with no galectin-1 IR, but their sizes were clearly decreased compared with SOD1G93A/Lgals1+/+ mice. Cell nuclei were counter-stained with DAPI (blue). Scale bars, 100 μm.

Figure S3. Quantitative Western blotting of galectin-1 in extracts prepared from whole spinal cord. Two additional sets of data for Figure 4C obtained from wild-type (WT) and SOD1G93A/Lgals1+/+ (SOD) mice as shown in Figure 4B are presented.

Figure S4. Kaplan–Meire graphs showing the onset of rotarod failure. (A) and 10% body weight loss (B). There was no statistically significant difference between SOD1G93A/Lgals1+/+ mice (n = 11) and SOD1G93A/Lgals1−/− mice (n = 11) in the onset of rotarod failure (P = 0.5451, log-rank test) and 10% body weight loss (P = 0.8201, log-rank test).

Figure S5. Electron microscopy of myelinated axons in the lumbar cord. An additional three sets of images from the electron microscopy data shown in Figure 6B are presented. Swollen axons (≥4 μm) were marked with an asterisk. Black asterisks show axons containing vacuoles and white asterisks show axons without vacuoles. The diameters of swollen axons in SOD1G93A/Lgals1−/− mice were smaller than those in SOD1G93A/Lgals1+/+ mice and accumulation of vacuoles was less frequently observed in SOD1G93A/Lgals1−/− mice, even in swollen axons. Scale bars, 5 μm.

Figure S6. Frequency distribution of axon diameters. Relative frequency (%) of axons with given diameter in wild-type, SOD1G93A/Lgals1+/+ and SOD1G93A/Lgals1−/− mice are shown in bar graphs. In SOD1G93A/Lgals1+/+ and SOD1G93A/Lgals1−/− mice, the relative frequencies of axons containing vacuoles are shown by dark bars. In SOD1G93A/Lgals1+/+ mice, axons with an internal diameter larger than 4 μm were increased and most of those contained vacuoles. In SOD1G93A/Lgals1−/− mice, large swollen axons were less frequently observed and the proportion of axons containing vacuoles was lower compared with SOD1G93A/Lgals1+/+ mice. Three mice were examined in each group, and the total numbers of axons examined were 318 axons in wild-type, 911 axons in SOD1G93A/Lgals1+/+ and 1071 axons in SOD1G93A/Lgals1−/−. Error bars show the mean ± SEM.


Video S1. Time-lapse imaging of CTB-labelled vesicles in isolated sciatic nerve from wild-type mice at 28 weeks of age. Images recorded for 174.4 s are shown.


Video S2. Time-lapse imaging of CTB-labelled vesicles in isolated sciatic nerve from SOD1G93A/Lgals1+/+ mice at 28 weeks of age. Images recorded for 174.4 s are shown.


Video S3. Time-lapse imaging of CTB-labelled vesicles in isolated sciatic nerve from SOD1G93A/Lgals1−/− mice at 28 weeks of age. Images recorded for 174.4 s are shown.

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