NGF increases the level of autophagy in nerve lesions at day 5 after injury. (A) Western blotting analysis of ATG-7, ATG-5, Beclin-1 and LC3 in sham, PNI and PNI+NGF groups at 5 days post crush. (B-E) Quantification of autophagy-related proteins expressed in (A). GAPDH was set as a loading control. Data are presented as the mean ± SEM; n = 3 or 4 independent experiments. ATG-7 F(2, 6) = 32.47, **Psham vs PNI = 0.006, *PPNI vs PNI+NGF = 0.036; ATG-5 F(2, 6) = 20.90, *Psham vs PNI = 0.032, *PPNI vs PNI+NGF = 0.041; Beclin-1 F(2, 6) = 29.82, *Psham vs PNI = 0.027, *PPNI vs PNI+NGF = 0.039; LC3II/I F(2, 9) =55.81, **Psham vs PNI = 0.0084, *P = 0.031. (F, G) Representative images of LC3 (green) immunostaining and quantitative analysis of the average LC3 positive area in each group. n = 3 rats per group, and the results are shown as the mean ± SEM. F(2, 6) = 61.36, **Psham vs PNI = 0.0072, *PPNI vs PNI+NGF = 0.041.