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. 2023 Apr 9;20(5):689–701. doi: 10.7150/ijms.76580

Figure 3.

Figure 3

Hypoxia and histone H3 activate NLRP3 inflammasome through TLR9 (A) The expression of HIF-1α, TLR9, NLRP3, Caspase-1, IL-1β and IL-18 were determined by western blot in Huh7 and G2 cells exposed to 24h hypoxia. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, N.S. no significance. (B) The expression of caspase-1 in Huh7 and G2 cells treated with 10 μg/ml anti-Histone H3 neutralizing antibody under hypoxia. (C) The IP results showed that the content of histone H3 bound by TLR9 increased after hypoxia. (D) The content of H3 in cytoplasm of Huh7 and G2 cells treated with 30 μg/ml recombinant human histone H3 for 24 hours. (E) The expression of caspase-1 in Huh7 and G2 cells treated with 30 μg/ml recombinant human histone H3 for diverse times. (F) The expression of TLR9, NLRP3, caspase-1, IL-1β and IL-18 were significantly increased in stable histones H3-expressing cells via western blot analysis. (G) Western blot analysis for caspase-1 from stable histones H3-expressing cells after transfected with TLR9 siRNA. Lv-control+si-control group was stable Lv-control HCC cell to be transfected with si-control; Lv-H3+si-TLR9 group was stable histones H3-expressing cells to be transfected with si-TLR9.