Fig. 2

Autophagy impairment in the TNC after chronic NTG administration. a–c Representative western blot of autophagy-related proteins following NTG injection. Western blot assay revealed increased ration of LC3II/LC3I (a), in combination with the up-regulation of p62 (b), without any change in beclin1 (c) under the chronic stimulation with NTG. n = 6 per group; data are presented as mean ± SEM; independent-sample t test; **p < 0.01, ***p < 0.001 compared with the SHAM group; to further determine the autophagic flux, mice were treated with CQ, an inhibitor of lysosome function, every day for 9 days prior to NTG/VEH administration. d–f The representative immunoblot showing the change of autophagy-related proteins after CQ treatment. Quantitative analysis of band intensities showed that, after CQ treatment, LC3II did not further increase (d), the expression of p62 tended to increase, but the difference was not statistically significant (e), and the level of beclin1 did not change (f), which confirmed the inhibition of autophagic flux in CM model. n = 6 per group; values are expressed as mean ± SEM; one-way ANOVA and Tukey’s multiple comparison test; ***p < 0.001 compared with the SHAM-VEH group