Fig. 1

Role of RIP3 in liver fibrosis induced by schistosomiasis. a Western blot detection of RIP3, RIP1, pMLKL and cleaved caspase 3 expression in the liver. b Western blot detection of RIP3 expression in CD45⁺ cells, HSCs, and hepatocytes. c Comparison of the survival rates of WT and RIP3^−/− mice before and after schistosome infection (n = 5). d Weight gain percentage of WT and RIP3^−/− mice before and after schistosome infection (n = 5). e Egg granuloma in infected WT and RIP3^−/− mouse livers at 0 w, 6 w, 8 w, and 12 w (HE staining, scale = 100 μm). f Serum ALT and AST levels of infected WT and RIP3^−/− mice at 0 w, 6 w, 8 w, and 12 w (n = 5). g Degree of liver fibrosis in infected WT and RIP3^−/− mice at 0 w, 6 w, 8 w, and 12 w (Masson staining, scale = 100 μm). Collagen was dyed blue. h Hydroxyproline levels in WT and RIP3^−/− mouse liver tissues at 0 w, 6 w, and 8 w (n = 5). i Collagen I and α-SMA expression levels in HSCs from infected or noninfected (control) mice detected by Western blot analysis. j The mRNA expression levels of TNF-α, IL-6, IL-1β, and MCP-1 in infected WT and RIP3^−/− mouse livers at 0 w, 6 w, 8 w, and 12 w detected by fluorescence quantitative PCR (n = 5). k Flow cytometry detection of the ratio of macrophages, CD3⁺ T cells, and CD45⁺ cells in mouse livers before and after infection. l TUNEL staining of infected mouse livers at 0 w, 6 w, 8 w, and 12 w. Dead cells were dyed green. Nuclei were stained with DAPI (blue) (scale = 200 μm). m RIP3, RIP1, pRIP1, and pMLKL expression levels in infected mouse livers. n ROS levels in infected WT and RIP3^−/− mouse livers at 0 w, 6 w, 8 w, and 12 w (n = 5). o Western blot detection of JNK, Erk1/2, P38, and phosphorylated protein expression in WT and RIP3^−/− mouse livers before and 8 w after infection. p Immunohistochemistry detection of p-cJUN expression (brown) in WT and RIP3^−/− mouse livers before and after infection with S. japonicum (scale = 50 μm). Nuclei were stained with hematoxylin (blue). q Immunohistochemistry detection of Egr1 expression (brown) in WT and RIP3^−/− mouse livers before and after infection with S. japonicum (scale = 50 μm). Nuclei were stained with hematoxylin (blue). r Expression of NF-κB, Nrf-2 and Egr1 in WT and RIP3^−/− mouse livers before and 8 weeks after infection, as detected by Western blot analysis. s Egg granuloma (HE staining, scale = 50 μm) and fibrosis (Masson staining, scale = 50 μm) in SP600125-treated infected mice (n = 5). Collagen was dyed blue by Masson staining. t Immunohistochemistry detection of α-SMA expression (brown) in the livers of SP600125-treated infected mice. Nuclei were stained with hematoxylin (blue) (scale = 50 μm). u TNF-α, F4/80, IL-1β, IL-6, and MCP-1 mRNA levels in the livers of SP600125-treated infected mice, as detected by fluorescence quantitative PCR (n = 5). v RIP3 expression in the livers of SP600125-treated infected mice, as detected by Western blot analysis. w Immunohistochemistry detection of Egr1 (brown) expression in liver (scale = 50 μm). Nuclei were stained with hematoxylin (blue). Experiments were repeated ≥3 times, and all the data are shown as the mean ± SD, *P < 0.05; **P < 0.01; ***P < 0.001