Fig. 3

Knockdown or inhibition of TLR9 activation reduced NPC pyroptosis under OS. Knockdown by siRNA or inhibition of TLR9 activation by E6446 with 100 µM H₂O₂-treated the NPCs for 24 h. a Agar gelatin electrophoresis images of TLR9 cDNA in the negative control, si-Scrambled, and si-TLR9 groups. b RT-qPCR quantitative analysis of TLR9 cDNA after siRNA knockdown in the negative control, si-Scrambled, and si-TLR9 groups. c Western blotting images of the expression levels of TLR9 and NF-κB after siRNA knockdown in the negative control, si-Scrambled, and si-TLR9 groups. d, e Quantitative analysis of the protein level of TLR9 and NF-κB after siRNA knockdown in the negative control, si-Scrambled, and si-TLR9 groups. f, g Immunofluorescence images of TLR9 and NF-κB in the negative control, si-Scrambled, and si-TLR9 groups. h Western blotting images of NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in the negative control, si-Scrambled, and si-TLR9 groups. i Fluorescence images of Calcein AM/PI staining in the negative control, si-Scrambled, and si-TLR9 groups. j TEM images of the NPCs in the negative control, si-Scrambled, and si-TLR9 groups. k Quantitative analysis of the expression levels of NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in the negative control, si-Scrambled, and si-TLR9 groups. l Quantitative analysis of IL-1β by ELISA in the negative control, si-Scrambled, and si-TLR9 groups. m Western blotting images of NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in the E6446- or PBS-treated NPCs. n Fluorescence images of Calcein AM/PI staining in the E6446- or PBS-treated NPCs. o TEM images of the NPCs in the E6446- or PBS-treated NPCs. p Quantitative analysis of the expression levels of NLRP3, ASC, pro caspase-1, cleaved caspase-1, GSDMD, and cleaved GSDMD in the E6446- or PBS-treated NPCs. q Quantitative analysis of IL-1β by ELISA in the E6446- or PBS-treated NPCs. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001