Quantifying Post-Injury Lesion Volume and CST Destruction

Post-injury lesion volume was quantified as previously described^{15 (link)}. Briefly, coronal brain sections were stained using Cresyl violet (Nissl stain; Sigma-Aldrich). The area of lesioned structures was expressed as a percentage of the volume of corresponding structures in the contralesional hemisphere. Lesion volume was measured using ImageJ (National Institutes of Health, Bethesda, MD, USA); the percentage of the lesion volume was calculated as (contralesional cortex–ipsilesional cortex)/contralesional cortex. To assess CST destruction volume, transverse cervical cord sections were stained using anti-PKCγ antibody. CST area was expressed as a percentage of the PKCγ-immunoreactive area of the uninjured CST (C4–C7). PKCγ staining intensity was measured using ImageJ; the percentage of the lesion volume was calculated as (uninjured CST–injured CST)/uninjured CST.