SOD1^G93A mice have a lower hind limb muscle mass than Ntg controls already at a presymptomatic stage of the disease.

<p>(A-B) Grip strength as latency to fall (sec) and body weights (g) in Ntg and SOD1^G93A mice are reported from 11 to 22 weeks of age (n = 10 per group). Data are expressed as mean ± SEM. *, p < 0.05 by two-way ANOVA for repeated measures (the interaction between genotype and time was significant in A and B with p < 0.0001), Bonferroni’s post-hoc test. (C-D) The weight of TA (C) and Gastrocnemius (GC) (D) was measured in Ntg and SOD1^G93A mice at 12, 14, 17 and 20 weeks of age (n = 5 for each group). The weight of TA and GC was lower in SOD1^G93A mice than age-matched Ntg controls already at a presymptomatic stage of the disease (12 weeks of age). Muscle weight was expressed in mg. Decrease in % was reported with respect to Ntg mice at 12 weeks of age. Data (mean ± SEM) were significantly different (p < 0.05), by one-way ANOVA, Newman-Keuls <i>post-hoc</i> test: *, <i>versus</i> Ntg; #, <i>versus</i> 12-week old SOD1^G93A; °, <i>versus</i> 14-week old SOD1^G93A;§, <i>versus</i> 17-week old SOD1^G93A mice.</p

Micro-CT evaluation of the hind limb muscle mass and definition of the index of muscle mass (IMM).

<p>(A) Mice were placed prone on the micro-CT bed leaving the hind limbs in a natural position (foot and tibia angle mean ± SD: 41° ± 6, n = 30). (B) Scanned raw images were reconstructed in 3D and analyzed using Micro View analysis software. Scale bar, 2 mm. (C) For the evaluation of the muscle mass, the following parameters from 3D reconstructed images were measured: the distance from the upper extremity of the tibia (ut) to the medial malleolus (m), defined as the length of the tibia (L), and the perpendicular distance from the half-length of the tibia to the external margin of the hind limb muscle, referred to as the thickness of the muscle (T). These measurements were assessed on the plane in which the patella (p) and the upper extremity of tibia (ut) were clearly visualized. Scale bar, 2 mm.</p

IMM correlates with hind limb muscle weight in SOD1^G93A mice as disease progresses.

<p>(A) IMM was evaluated in Ntg and longitudinally in SOD1^G93A mice at 12, 14, 17 and 20 weeks of age (n = 5 for each group). Decrease in % was reported respect to Ntg mice at 12 weeks of age. Data, expressed as mean ± SEM, were statistically significant different (p < 0.05), by one-way ANOVA, Newman-Keuls <i>post-hoc</i> test: *, <i>versus</i> Ntg; #, <i>versus</i> 12-week old SOD1^G93A mice. (B-C) A correlation analysis was done with the mean values of the IMM and of the muscle weight of TA (B) and Gastrocnemius (GC) (C) for the different experimental groups. The analysis confirmed a significant (p < 0.008 for TA, p < 0.005 for GC) direct correlation between the two type of measurements with r² = 0.9293 and r² = 0.9502 for TA and GC respectively. Data are expressed as mean ± SEM.</p

Additional file 3: of RNS60 exerts therapeutic effects in the SOD1 ALS mouse model through protective glia and peripheral nerve rescue

Figure S3. A-D) Representative images of LSC micrographs stained with S100β (green) and GFAP (red) at 20 weeks of age. Scale bar: 20 μm. E) Quantification of immunofluorescence showed no differences between the two transgenic groups. Bar graphs represents mean ± SEM, (n = 5 animals per group); One-way ANOVA followed by post hoc Fisher’s LSD non-parametric test (p = 0.420). (DOCX 1342 kb

Additional file 2: of RNS60 exerts therapeutic effects in the SOD1 ALS mouse model through protective glia and peripheral nerve rescue

Figure S2. Representative immunoblot for GFAP performed on ventral portion of LSC of NTG mice or transgenic mice treated with NS or RNS60, at 20 weeks of age, and relative quantification. Data are expressed as mean ± SEM, (n = 5 animals per group). Data were statistically analyzed using one way ANOVA followed by post hoc Fisher’s LSD. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, n.s. = non significant. (DOCX 162 kb