Xenografts for LOXIMVI sgEGFP (WT) and LOXIMVI sgGPX4 (KO) cells were established by injecting 10 million cells in a 1:1 PBS:Matrigel mixture containing 2.5 uM ferrostatin-1 into the flanks of athymic mice (NRC Nude, Taconic). Animals were dosed daily with 2 mg kg-1 ferrostatin-1 by intraperitoneal injections. Tumour volume was measured twice a week.

Eight week-old male nude mice weighing approximately 21-23 g were divided into two groups (DMSO group and erastin group) with 5 mice per group. Each mouse was injected with 5 x 10⁶ cells transfected non-targeting shRNA and idh2-targeting shRNA on the left and right hind legs, respectively. Erastin was administered intraperitoneally at 5 mg/kg for 15 consecutive days starting on the same day as tumor injection.

For the in vivo xenograft mouse model, 17-b-estradiol 60-d release pellets (Innovative Research of America) were implanted subcutaneously into the left flank 7 d before tumor inoculation. GPX4 iKO BT474 cells were inoculated by injecting 5 x 10⁶ cells in 50% Matrigel subcutaneously in the right flank of 6- to 8-wk-old female athymicnu/numice (Envigo). For PC-3 tumor models, male athymic nu/nu mice aged 5 to 6 wk were injected in the right flank with 5 x 10⁶ PC-3 cells.

Adult male Sprague-Dawley rats (SD rats, weighing 250-300 g) aged 11-12 weeks were purchased from SLAC Laboratory Animal Co., Ltd. (Shanghai, China). All 96 rats were randomly divided into four groups of 24 rats each: Sham group, Sham + IRN (30 mg/Kg) group, ICH group, and ICH + IRN (30 mg/Kg) group. The rats in sham group were injected with PBS solution, and the Sham + IRN (30 mg/Kg) group was received an equal amount of 30 mg/Kg IRN solution (intra-peritoneal injection) after the sham operation. After ICH, the rats in ICH group were injected with PBS solution, and the ICH + IRN (30 mg/Kg) group was received an equal amount of 30 mg/Kg IRN solution (intra-peritoneal injection).

Six- to eight-week-old female NSG or C57BL/6 mice were obtained from the Jackson Laboratory. For HT-1080 tumor model, 10⁶ tumor cells were subcutaneously injected on the right flank of NSG mice. For adoptive transfer of OT-I to B16-OVA model, 105 B16-OVA cells were subcutaneously injected on the right flank of C57BL/6 mice. For the B16 tumor model, 105 B16F0 cells were subcutaneously injected on the right flank of C57BL/6 mice. For the ID8 tumor model, 2 x 10⁶ luciferase-expressing ID8 cells were injected into the peritoneal cavity of each female mouse.

Six- to eight-week-old female NSG or C57BL/6 mice were obtained from the Jackson Laboratory. For HT-1080 tumor model, 10⁶ tumor cells were subcutaneously injected on the right flank of NSG mice. For adoptive transfer of OT-I to B16-OVA model, 105 B16-OVA cells were subcutaneously injected on the right flank of C57BL/6 mice. For the B16 tumor model, 105 B16F0 cells were subcutaneously injected on the right flank of C57BL/6 mice. For the ID8 tumor model, 2 x 10⁶ luciferase-expressing ID8 cells were injected into the peritoneal cavity of each female mouse.