ROS-induced lysosomal dysfuntion initiates autophagy flux blockade and ER stress-induced apoptosis after SCI. (A) ELISA of oxidation products 8-OHdG, AOPP, and MDA in spinal cord lesions from Control and SCI mice at the indicated time points. (B) ELISA of 8-OHdG, AOPP, and MDA in spinal cord lesions from mice grouped as indicated at Day1 after SCI. (C) Western blotting of Beclin1, VPS34, C-CTSD, SQSTM1/p62, UB and LC3 in spinal cord from non-SCI (Control) mice and SCI mice treated with MnTBAP or Vehicle1 at Day1 after SCI. (D) Western blot analysis of LC3 in SCI+Vehicle1, and SCI treated with MnTBAP spinal cord slides at Day1 cultured in the presence or absence of CQ. (E) Densitometric analysis of Beclin1, VPS34, ATP6V1B2, LAMP2, C-CTSD, SQSTM1/p62, UB and LC3II from (C) normalized to loading control GAPDH. (F) Densitometric analysis of LC3II from (D) normalized to the loading control GAPDH. (G) Western blot analysis of GRP78, PDI, PERK, p-PERK, eIF2α, p-eIF2α, ATF4, CHOP, CASP12, C-CASP12, and C-CASP3 in spinal cord lesions from the grouped mice on Day3. (H) Densitometric analysis of GRP78, PDI, p-PERK, p-eIF2α, ATF4, CHOP, C-CASP12 and C-CASP3 from (G) normalized to loading control GAPDH. n=6, ns stands for not significant, *P<0.05, **P<0.01.