Fig. 4.

Effect of iron ions on CCCP-induced pyroptosis of B cells. a Interference of splenic B cells of mice was induced using FeSO₄ and CCCP. After 24 h, the morphology of the B cells was observed. The red arrow indicates swollen B cells. b Detection of B cell viability by CCK8. c Detection of LDH release from B cell supernatants using an LDH kit. d Detection of the levels of MCP-1 in B cell supernatants by ELISA. e Detection of the expression levels of pyroptosis-related proteins in B cells by western blot. f After Tom20-KD via siRNA transfection, the expression of Tom20 was detected. g The morphology of B cells was observed after 24 h. The red arrow indicates swollen B cells. h Detection of B cell viability by CCK8 after TOM20-KD. i Detection of LDH release in B cell supernatants after Tom20-KD using an LDH kit. j Detection of the levels of MCP-1 in B cell supernatants after Tom20-KD via ELISA. k Western blot detection of the expression of pyroptosis-related proteins in B cells after Tom20-KD. l Morphological and immunofluorescence identification of spinal cord neurons in mice; NeuN was red. m–p After intervention, the supernatant of B cells was co-cultured with spinal cord neurons, and the apoptosis of neurons was observed by TUNEL staining. TUNEL was green, and dapi was blue. All data are expressed as the mean ± SD (n ≥ 3 replicates per group). ns P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001. CCCP carbonyl cyanide m-chlorophenyl hydrazone, CCK8 cell counting kit 8, LDH lactate dehydrogenase, MCP-1 monocyte chemoattractant protein 1, ELISA enzyme-linked immunosorbent assay, Tom20-KD Tom20 knock-down, siRNA small interfering ribonucleic acid, SD standard deviation, ns not significant