Nogo a

Sections were incubated for 1 h in blocking solution containing PBS 0.1% Triton X-100 (v/v) and 5% serum (v/v) and incubated overnight in the primary antibody in 2% blocking solution. The following antibodies were used: ubiquitin (Dako), Nogo-A (R&D Systems), myosin heavy chain subtypes to identify the combination of muscle fibre type (Developmental Studies Hybridoma Bank: myosin heavy chain 2A BF-F3-s, or myosin heavy chain 2B SC-71-s) (see Table S1 for full details of primary antibodies). Primary antibodies were detected using Alexa Fluor 488 or Alexa Fluor 568 secondary antibodies (see Table S1 for full details) and nuclei were stained with DAPI (Sigma-Aldrich). Images were acquired using a Leica DMR fluorescence microscope.
EDL was stained with α-Bungarotoxin (Sigma-Aldrich, T0195) which labels the postsynaptic acetyl choline receptor, and then stained with ChAT antibody (Millipore), a marker of motor axons (see Table S1). Analysis of innervation was performed as follows: percentage of end plates fully innervated; percentage of end plates partially innervated; percentage of end plates completely denervated.