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. 2024 Oct 14;14:23988. doi: 10.1038/s41598-024-74742-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 5 — Differential protective effect of catechol estrogens, their parent hormones and the monomethoxy estrogen metabolites against RSL3-induced cell death in HT22 cells. (A–B) Effect of RSL3 on cell viability (MTT assay; (A)) and gross cell morphology (B). Cells were treated for 24 h with different concentrations of RSL3 (A), or treated with 80 nM RSL3 for different durations (B). Each value is the mean ± S.D. (n = 5). (C) Protective effect of Fer-1 against RSL3-induced cell death (MTT assay, n = 4). Cells were treated for 24 h with RSL3 (80 nM) ± Fer-1 (1 μM). Note that in this experiment (conducted separately at a different time), the cells experienced a reduced sensitivity to RSL3 cytotoxicity. (D–G) Cells were treated with RSL3 (80 nM) ± a catechol estrogen (2-OH-E₁, 2-OH-E₂, 4-OH-E₁ or 4-OH-E₂) at indicated concentrations for 24 h, and cell viability was determined by MTT assay. Each value is the mean ± S.D. (n = 5). (H, I) Comparison of the protective effect of E₁ and E₂ against RSL3-induced cytotoxicity (MTT assay). Cells were treated with different concentrations of E₁ (H) or E₂ (I) for 24 h. Each value is the mean ± S.D. (n = 5). (J–M) Comparison of the protective effect of the monomethoxy estrogen metabolites against RSL3-induced cytotoxicity (MTT assay). Cells were treated with different concentrations of the estrogen metabolites for 24 h. Each value is the mean ± S.D. (n = 5). (N) Protective effect of cystamine against RSL3-induced ferroptotic cell death. The cell viability was determined by MTT assay. Each value is the mean ± S.D. (n = 5). (O) Protective effect of CPZ and SMT against RSL3-induced cell death (MTT assay, n = 5). Cells were treated for 24 h with RSL3 (80 nM) ± CPZ (20 μM) or with RSL3 (80 nM) ± SMT (100 μM). Note that in this experiment (which conducted separately at a different time), the cells experienced a reduced sensitivity to erastin-induced cytotoxicity. The quantitative data are presented as mean ± S.D. * or ^#, P < 0.05; ** or ^##, P < 0.01. n.s., not significant. (* or ^#denote statistical significance in comparison with the control group or the RSL3 alone group, respectively).