Fig. 5

Inhibition of mPTP opening blocked cytosolic mtDNA release and reduced NPC pyroptosis. CsA or DMSO + 100 µM H₂O₂ was used to treat NPCs for 24 h. a, b Fluorescence images of Calcein AM/Co2⁺ staining and quantitative analysis of fluorescence intensity for the opening of mPTPs in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. c, d Fluorescence images of Mito-tracker and quantitative analysis of fluorescence intensity for mitochondrial activity in the CsA- or DMSO-treated NPCs. e Fluorescence images of dsDNA (green) in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. f The PLA immunofluorescence images of mtDNA-TLR9 interaction in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. g, h Immunoprecipitation images of mtDNA-TLR9 interaction and quantitative analysis of the level of mtDNA-TLR9 interaction in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. i Agar gelatin electrophoresis images of mtDNA (ND1 and ND2) in the cytoplasmic or total lysates in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. j Quantitative analysis of the level of mtDNA (ND1 and ND2) in the cytoplasm in the CsA or DMSO + 100 µM H₂O₂ treated NPCs. k, l Western blotting images and quantitative analysis of the expression levels of TLR9 and NF-κB in the CsA- or DMSO-treated NPCs. m, n Immunofluorescence images of TLR9 and NF-κB in the CsA- or DMSO-treated NPCs. o Western blotting images of NLRP3, ASC, pro caspase-1, cleaved caspase − 1, GSDMD, and cleaved GSDMD in the CsA- or DMSO-treated NPCs. p Quantitative analysis of the expression levels of NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in the CsA- or DMSO-treated NPCs. q Quantitative analysis of IL-1β by ELISA in the CsA- or DMSO-treated NPCs. r TEM images of the NPCs in the CsA- or DMSO-treated. s Fluorescence images of Calcein AM/PI staining in the CsA- or DMSO-treated NPCs. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001