Ferroptosis is suppressed in liver cancer. (A) TMA of c-PARP, HMGB1 and GPX4 in liver cancer and normal liver tissues, as measured by IHC. (B–C) Western blots and IHC of c-PARP, HMGB1 and GPX4 in fresh normal liver and liver cancer tissues (B), and established hepatocyte and liver cancer cell lines, as indicated (C). The IHC images in Fig. 1B are representative ones from patient #1. Scale bar, 200 μm. (D) Ferroptosis inhibited liver tumorigenesis in DEN/CCl4-treated mice. Erastin (10 mg/kg) with or without ferrostatin-1 (2 mg/kg) was treated after tumors formed in mice. The liver-bearing tumor in mice with the indicated treatment was shown. Scale bar, 1 cm, n = 5/group. The expressions of c-PARP, HMGB1 and GPX4 in the tumor were measured by IHC. Scale bar, 200 μm n = 5/group. (E) Stimulation of the ferroptosis-inhibited formation of xenografts. Xenografts in mice with indicated treatments are shown. Scale bar, 1 cm, n = 5/group. The expression of c-PARP, HMGB1 and GPX4 in xenografts were measured by IHC. Scale bar, 200 μm n = 5/group. Images of IHC and WB are representative ones of 3–5 independent experiments (except Fig. 1A).