HSC-2 or K19-Wnt1/C2mE-KP cells (2 x 10⁶ cells per site) were implanted subcutaneously in the flank of athymic nude mice (CLEA Japan) or C57BL6 mice (CLEA Japan), respectively. The mice were then injected intraperitoneally with physiological saline or sulfasalazine (350 mg/kg per day), or with combinations of physiological saline, sulfasalazine (400 mg/kg per day), and dyclonine hydrochloride (5 mg/kg per day).

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN9 cells with transfection of CDH1 or control vector or HN4 cells with ZEB1 or control vector were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route). Each group included six mice.

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN9 cells with transfection of CDH1 or control vector or HN4 cells with ZEB1 or control vector were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route). Each group included six mice.

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN9 cells with transfection of CDH1 or control vector or HN4 cells with ZEB1 or control vector were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route). Each group included six mice.

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from Central Lab Animal Inc. (Seoul, Republic of Korea). HN9 cells with shDLD or vector control were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route).

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN4 parental cells with transfection of vector control or shGPX4 were subcutaneously injected into the bilateral flank of nude mice. The same was performed in HN4 erPCC with transfection of control vector or shGPX4. Each group included six mice. In other experiments, HN4 parental cells with transfection of control vector or shMPC1 were injected to nude mice in the same way above. HN4 parental cells and erPCC without target gene silencing were also injected into nude mice. Each group included six mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route).

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN4 parental cells with transfection of vector control or shGPX4 were subcutaneously injected into the bilateral flank of nude mice. The same was performed in HN4 erPCC with transfection of control vector or shGPX4. Each group included six mice. In other experiments, HN4 parental cells with transfection of control vector or shMPC1 were injected to nude mice in the same way above. HN4 parental cells and erPCC without target gene silencing were also injected into nude mice. Each group included six mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route).

Five-week-old athymic BALB/c male nude mice (nu/nu) were purchased from Central Lab Animal Inc. (Seoul, Republic of Korea). HN4R cells with vector control or shGLRX5 were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or SAS (250 mg/kg daily per intraperitoneal route). Each group included seven mice.

Five-week-old athymic BALB/c male nude mice (nu/nu) were purchased from Central Lab Animal Inc. (Seoul, Republic of Korea). HN4R cells with vector control or shGLRX5 were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or SAS (250 mg/kg daily per intraperitoneal route). Each group included seven mice.

Five-week-old athymic BALB/c male nude mice (nu/nu) were purchased from Central Lab Animal Inc. (Seoul, Republic of Korea). HN4R cells with vector control or shGLRX5 were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or SAS (250 mg/kg daily per intraperitoneal route). Each group included seven mice.

Five-week-old athymic BALB/c male nude mice (nu/nu) were purchased from Central Lab Animal Inc. (Seoul, Republic of Korea). HN4R cells with vector control or shGLRX5 were subcutaneously injected into the bilateral flank of nude mice. From the day when gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or SAS (250 mg/kg daily per intraperitoneal route). Each group included seven mice.

Six-week-old athymic BALB/c male nude mice (nu/nu) were purchased from OrientBio (Seoul, Republic of Korea). HN4 cells with transfection of PGRMC1 overexpression or control vector and HN4PCC with shPGRMC1 or control vector were subcutaneously injected into the bilateral flank of nude mice. When gross nodules were detected in tumor implants, mice were subjected to different treatments: vehicle or sulfasalazine (250 mg/kg daily per intraperitoneal route).

Four-week-old male BALB/c-nu mice were purchased from the Shanghai Laboratory Animal Center (Shanghai, China). About 4 x 10⁶ CAL27 cells were stably transfected with lentivirus. After administration of 2 ug/mL puromycin for three days, transfection efficiency was confirmed by western blotting. Approximately 2 x 10⁶ transfected cells were subcutaneously injected into flanks. For the drug-administration study, 20 mg/kg erastin (S7242, Selleck Chemicals) were administrated intraperitoneally twice every other day. Approximately 20 uL IL-6 (10 ug/mL, PeproTech, USA) were given intratumorally twice every other day.

Four-week-old male BALB/c-nu mice were purchased from the Shanghai Laboratory Animal Center (Shanghai, China). About 4 x 10⁶ CAL27 cells were stably transfected with lentivirus. After administration of 2 ug/mL puromycin for three days, transfection efficiency was confirmed by western blotting. Approximately 2 x 10⁶ transfected cells were subcutaneously injected into flanks. For the drug-administration study, 20 mg/kg erastin (S7242, Selleck Chemicals) were administrated intraperitoneally twice every other day. Approximately 20 uL IL-6 (10 ug/mL, PeproTech, USA) were given intratumorally twice every other day.

Six-week-old athymic BALB/c malenude mice(nu/nu) were purchased from the Central Lab Animal Inc. (Seoul, Republic of Korea). SAS-resistant HN10 cells were injected subcutaneously into the flank of nude mice. Following the detection of gross nodules from the tumor implants, the mice were subjected to four different treatments: vehicle, SAS (250mg/kg daily per intraperitoneal route), PGZ (20 mg/kg daily per oral administration), or SAS plus PGZ.