Fig. 5: Ferroptosis and lipid peroxidation are induced in mammary tumor-bearing mice.

a Female BALB/c mice carrying the 4T1 cell-formed tumors at the mammary fat pads were treated with RSL3 alone or combined with rosiglitazone (Rosi). Tumor growth in mice was presented by images (b), as well as assessed by tumor weight (c, ^***P < 0.001 vs the control group and ^##P < 0.01 vs the RSL3 group, by independent samples t-test, respectively) and tumor volume (d, ^***P < 0.001 vs the control group and ^###P < 0.001 vs the RSL3 group, by two-way repeated measures AVONA). Data are mean ± SEM (n = 5 independent biologically animals). e TUNEL assay was carried out to evaluate the death ratio of tumor cells. Data are mean ± SEM (n = 3 independent biologically animals). f Tumor cells were measured by MDA assay. Data are mean ± SEM (n = 3 independent biologically animals). ^**P < 0.01 and ^***P < 0.001, by one-way ANOVA with LSD post hoc test. g Immunohistochemistry of tumor sections labeled with antibodies of 4-HNE and GPX4. Scale = 50 μM. h Western blotting analysis of 4-HNE and GPX4 expression in tumor. Each immunoblot represents three biological repeats, and representative blotting results are shown. i Content of SAPE-OOH in tumor cells was determined by LC–MS/MS. Data are mean ± SEM (n = 3 independent biologically animals). ^*P < 0.05, ^**P < 0.01 and ^***P < 0.001 by one-way ANOVA with LSD post hoc test. All data represent mean ± SEM.