Validation of the lesion, levels of mTOR activity and CSPG degradation in pyramidotomy model. A, Representative coronal sections of PTEN^f/f;Rosa-tdTomato motor cortex stained with p-S6 and NeuN antibodies show depletion of p-S6 immunoreactivity in cortical neurons following rapamycin treatment. Rapamycin or vehicle was administered every third day, starting from the day of pyramidotomy and animals were perfused 4 weeks after injury. B, Representative coronal section of cervical spinal cord showing absence of PKC-γ immunoreactivity in the transected CST tract (n = 8). B′, Higher-magnification of the boxed area in B. C, C-4-S immunostaining to validate CSPG digestion in the cervical spinal cord (n = 3). Scale bars: A, 25 μm; B, C, 500 μm.