LPS/D-GalN-induced ALF enhances apoptosis and ferroptosis. (A) The survival rate of WT mice treated with vehicle or LPS/D-GalN (LPS, 20 μg/kg; D-GalN, 700 mg/kg) (n = 10). (B) Representative pictures of WT mice treated with vehicle or LPS/D-GalN for 3, 5, and 6 hours (n = 10). (C) Serum levels of ALT and AST with or without LPS/D-GalN co-injection (n = 6). (D) H&E and TUNEL staining after LPS/D-GalN co-injection (n = 6). (E) Immunohistochemistry of TNFα and F4/80 staining after LPS/D-GalN co-injection (n = 6). (F) Transmission electron microscope of liver tissues (n = 3). (G) MDA assay of liver homogenates (n = 5). (H and J) Western blot analyses of TFR, DMT1, GPX4, and XCT from WT mice (H) with or without LPS/D-GalN and (I) quantitative results (n = 3–9). (I and K) FSP1, DHODH, and POR proteins. Western blot analyses of WT mice (I) with or without LPS/D-GalN and (K) quantitative results (n = 3–9). (L) Immunohistochemistry of DMT1 staining and immunofluorescence of Ptgs2 in livers with or without LPS/D-GalN (n = 6). (M) Quantitative results of DMT1 and Ptgs2. All data were obtained from WT mice. Scale bars: 100 μm. Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. DAPI, 4′,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.