Autophagy evaluation in Schwann cells.

Representative pictures of LC3 (red) expression in SCs (GFAP–green) 7 days after CCI, in WT (A) or in Ambra1 (B) mice both in ST and CR condition; scale bar = 60 micron. (C) Western blot (WB) analysis for autophagic proteins (p-Atg13, p62 and LC3II) and quantification in the graph of LC3-II protein in ipsilateral ST vs ipsilateral CR nerves as ratio between LC3-II/actin proteins (P D) Sample images of SCs (GFAP–green) expressing p-mTOR (red) or p-AMPK (red) 3 days after ligature in WT mice in ST diet or CR regimen. (E) Proteins, extracted from naïve or ipsilateral sciatic nerves from WT and A+/- mice at 24h, 3 and 7 days from ligature in ST or RC condition, were analysed by WB using anti-pAMPK (T172), anti-AMPK, anti-p-mTOR (S2448), anti-mTOR. Both Actin and Hsp90 were used as loading controls. Densitometric analysis of p-AMPK over Actin is also shown. *P <0.05, **P<0.01, ***<0.0001 vs naïve).