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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) Representative analysis of BODIPY 581/591 C11 fluorescence at 540 nm emission wavelength (oxidized form, BODIPY-C11^Ox). Histograms show normalized cell counts in WT and Ninj1^−/− BMDMs left untreated or after treatment with 1 mM CuOOH for 30, 60, or 120 min. (B) Quantification of (A) with additional replicates included. Percentage of WT and Ninj1^−/− BMDMs positive for BODIPY-C11^Ox in untreated cells (UT) or after treatment with 1 mM CuOOH for 30, 60, or 120 min. (C) Percentage of WT and Ninj1^−/− BMDMs positive for BODIPY-C11^Ox in untreated cells (UT) or after treatment with 5 μM RSL3 for 30, 60, or 120 min. (D) Percentage of WT and Ninj1^−/− BMDMs positive for BODIPY-C11^Ox in untreated cells (UT) or after treatment with 5 μM ML162 for 30, 60, or 120 min. (E) Time course analysis from 0–4 h of Fluo-8 (intracellular Ca²⁺ probe) fluorescence signal in WT and Ninj1^−/− BMDMs treated with 1 mM CuOOH. When indicated, 25 μM Fer-1 was added simultaneously with ferroptosis activators (A–E). Data information: Graphs show the mean ± SD. Histograms show cell counts normalized to mode. Data are representative of three independent experiments performed in triplicate (A, E) or pooled from three independent experiments performed in triplicate (B–D). Source data are available online for this figure.