Fig. 4: Expression of the downstream necroptosis mediators RIPK3 and MLKL correlates with inhibition of autophagy and lysosomal function after SCI.

a The time course of RIPK3 and MLKL protein expression in spinal cord tissue surrounding injury site following SCI in mice. Full unedited western blots are presented in Supplemental Figure S5. b Quantification of RIPK3 data from a. c Quantification of MLKL data from a. n = 4. d IHC staining demonstrates accumulation of RIPK3 and p62/SQSTM1 in the same ventral horn cells at 24 h after SCI in mice. Images were acquired at ×20 magnification. Full time course images for RIPK3 are presented in Supplemental Figure S4b. e Quantification of RIPK3 data in d and S4b. n = 4. f Quantification of RIPK3 and SQSTM1 data in d. At day 1 after SCI 78.2% of RIPK3 positive cells were also positive for p62/SQSTM1. n = 12–17. g IHC staining demonstrates accumulation of MLKL and p62/SQSTM1 in the same ventral horn cells at 24 h after SCI in mice. Full time course images for MLKL are presented in Supplemental Figure S4c. h Quantification of MLKL data in g and S4c. n = 4. i Quantification of MLKL and SQSTM1 data in g. At day 1 after SCI 69.4% of MLKL positive cells were also positive for p62/SQSTM1. n = 10–12. All data are presented as mean±SE. *p < 0.05, **p < 0.01, ***p < 0.001