Apoptosis was increased during LPS/D-GalN-induced ALF. (A) LY6G immunohistochemistry staining of WT mice with or without LPS/D-GalN treatment. Scale bars: 100 μm. (B) Western blot analyses of caspase 9, BCL2, BAX, caspase 8, and cl-caspase 8 (cleaved-caspase 8) proteins from WT mouse liver treated with or without LPS/D-GalN co-injection (n = 5–6). (C) Quantitative results of caspase 9, BCL2, BAX, caspase 8, and cl-caspase 8. (D) Western blot analyses of RIP3 and GSDMD from WT mouse liver treated with or without LPS/D-GalN co-injection. (E) ratios of each protein to GAPDH (n = 3). (F) Ki67 immunofluorescence staining of L02 with or without LPS/D-GalN (LPS, 10 ng/mL; D-GalN, 1 mmol/L). (G) Cell Counting Kit-8 (CCK8) assay of LPS/D-GalN induced L02 injury pretreated with Fer-1 (1 μmol/L), ZVAD-FMK (10 μmol/L), and Nec-1 (10 μmol/L) (n = 5). Scale bar: 10 μm. Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. C, Control; F, Fer-1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; N, Necrostatin-1; Z, ZVAD-FMK.