Fig. 5. Histological evidence of improved muscular recovery by multiple episodes of distal nerve stimulation 6 weeks after nerve injury.

a Stained slices show no significant difference in mature axon (Tuj1, green color) signals in the regenerated nerve within the nerve conduit, indicating that the beneficial effect of motor recovery is not via accelerated axon regeneration. n = 3 repeated independently with similar results. b Immunohistochemical staining of the muscle fiber boundary (laminin, red color) reveals an increased muscle fiber surface area in the group with multiple episodes of distal nerve stimulation. n = 4 biologically independent animals. c Double staining of the neuromuscular junction demonstrates significantly increased overlapping of pre- (NFM, red color) and postsynaptic (alpha-bungarotoxin, green color) staining for the group with multiple episodes of distal nerve stimulation, indicating an enhanced number of neuromuscular junction (NMJ) and muscle reinnervation. Naive indicates mice with uninjured nerve. n = 4 biologically independent animals. The boxplots show the median (center line), the third and first quartiles (upper and lower edge of the box, respectively), and the largest and smallest value that is ≤1.5 times the interquartile range (the limits of the upper and lower whiskers, respectively). Statistical software (Version 6.0) was used for the analysis followed by a t-test and one-way ANOVA with Tukey multiple comparison analysis (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). Data available in source data file.