Figure 4. Rescue of motor function and death of motor neurons after long-term administration of FN1040 and FN58.

(A) Initial rotarod scores were not significantly different among the FN1040-treated (n = 13), the FN58-treated (n = 13), and the vehicle-treated (n = 13) AR2 mice. Mean ± SEM, one-way ANOVA followed by Tukey’s multiple-comparisons test. (B) Mice received either FN1040 or FN58 were able to retain on the rotarod longer than the vehicle-treated mice. FN58-pre: administration of initial dose of FN58 (5 μM). Mean ± SEM, two-way ANOVA followed by Tukey’s multiple-comparisons test, *P < 0.05, **P < 0.01 versus vehicle. ^#P < 0.05 versus initial value (Table S5). (C) Mice treated with FN1040 exhibited body weight change similar to those treated with vehicle, whereas those treated with FN58 failed to increase the body weight 1 mo after administration and thereafter. Mean ± SEM, two-way ANOVA followed by Tukey’s multiple-comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus vehicle (Table S6). (D, E, F, G) show morphological change. (D) Anterior horns of the cervical spinal cord double stained with anti-ChAT (red) and anti-TDP-43 (green). Scale bar = 20 μm. (E, F) show the number and the size of ChAT-positive cells, respectively, in the anterior horn of the cervical spinal cord from FN1040- or FN58-treated AR2 mice (n = 13), as compared with those in the untreated control AR2 mice (n = 13), vehicle-treated AR2 mice (n = 13) or C57BL4 (wt) mice (n = 12). 10 anterior horn cells per mouse were used for cell counting. Mean ± SEM, two-way ANOVA followed by Tukey’s multiple-comparisons test. ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (G) The proportion of the four TDP-43 localization patters in the ChAT-positive cells. Mean ± SEM, two-way ANOVA followed by Tukey’s multiple-comparisons test, compared with the nontreated mice (bars and asterisks in blue color and in red color are for the comparison for Nuc/Cyto and absent, respectively) (Table S7). Total number of MNs used for TDP-43 localization analysis; C57BL6 (n = 1,702), nontreated (n = 1,055), vehicle (n = 1,146), FN1040 (n = 1,575), and FN58 (n = 1,529).

Source data are available online for this figure.