Fig. 2

OS induced activation of the TLR9-NF-κB-NLRP3 axis and NPC pyroptosis. Human NPCs was cultured with 0, 100, and 200 µM H₂O₂ for 24 h. a Scatter plots of H2DCFDA staining for cytoplasmic ROS. b Quantitative analysis of H2DCFDA staining for cytoplasmic ROS. c Fluorescence images of MitoSOX for mitochondrial ROS. d Quantitative analysis of fluorescence intensity of MitoSOX for mitochondria ROS. e Immunoprecipitation images of mtDNA-TLR9 interaction. f Quantitative analysis of the level of mtDNA-TLR9 interaction. g The PLA immunofluorescence images of mtDNA-TLR9 interaction. h Immunofluorescence images of TLR9 in NPCs. i Immunofluorescence images of NF-κB in NPCs. j Western blotting images of TLR9, NF-κB, NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in NPCs. k Quantitative analysis of the expression levels of TLR9, NF-κB, NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in NPCs. l Fluorescence images of Calcein-AM/PI staining in NPCs. m TEM images of the NPCs. n Quantitative analysis of IL-1β in NPCs by ELISA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001