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. 2024 Feb 23;43(7):1164–1186. doi: 10.1038/s44318-024-00055-y

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, 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 changes were made. 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/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Western blot analysis of NINJ1 expression in (sgluc) and Ninj1 knockout (sgNinj1) NIH/3T3. (B) LDH release in WT (sgluc) and Ninj1 knockout (sgNinj1) NIH/3T3 after treatment with 1 mM CuOOH for 5 h, 5 μM RSL3 for 8 h, or 5 μM ML162 for 8 h. (C) Western blot analysis of NINJ1 expression in WT (crRNA ctrl) and Ninj1 knockout (crRNA Ninj1) MEFs. (D) LDH release in WT (crRNA ctrl) and Ninj1 knockout (crRNA Ninj1) MEFs after treatment with 0.4 mM CuOOH for 5 h, 5 μM RSL3 for 8 h, or ML162 5 μM for 8 h. (E) Western blot analysis of NINJ1 expression in WT (sgluc) and Ninj1 knockout (sgNinj1) RAW 264.7. (F) LDH release in WT (sgluc) and Ninj1 knockout (sgNinj1) RAW 264.7 after treatment with 1 mM CuOOH for 2 h, 5 μM RSL3 for 3 h, or 5 μM ML162 for 2 h. Cells were lentivirally transduced with Cas9 and a sgRNA targeting luciferase (sgluc) or crRNA (crRNA ctrl), used as controls, or NINJ1 (sgNinj1 and crRNA Ninj1). Cell extracts were analyzed (A, C, E). Vinculin (VIN), Tubulin (TUB), or GAPDH are loading controls. When indicated, 25 μM Fer-1 was added simultaneously with ferroptosis activators (B, D, F). Data information: All graphs show the mean ± SD. Data are representative of three independent experiments performed in triplicate. Source data are available online for this figure.