Nerve samples were prepared following a similar protocol. After methanol dehydration and peroxide bleaching, rehydrated samples were blocked at room temperature for 12 h in 10% goat serum (Sigma, St. Louis, Missouri) and 0.2% Triton-100 in PBS. Sciatic nerves were costained with mouse anti-βIII-Tubulin (801201, BioLegend, San Diego, California, 1:33 dilution) and rabbit SCG10 (Stathmin-2) (NBP149461, Novus Biologicals, Oakville, Canada, 1:200 dilution), in nerve wash solution (3% goat serum, 0.2% Triton-X, heparin in PBS) for 5 to 7 days at 37°C. After extensive washing, samples were incubated with Alexa 488 conjugated antimouse IgG (Invitrogen, Eugene, Oregon) and Alexa 568 conjugated antirabbit IgG, at a dilution of 1:33 and 1:200, respectively, for 5 to 7 days at 37°C. Finally, nerves were washed for 12 h in nerve wash solution and 30 min in PBS before storage at 4°C until clearing.