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. 2025 Oct 21;11:117. doi: 10.1038/s41540-025-00590-2

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© The Author(s) 2025

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/.

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Fig. 2 — A–C Average JNK C/N ratios of cells treated with sustained (A), transient (B), or pulsed (C) anisomycin. Treated cells are shown in red versus the DMSO vehicle control in black. Thick lines represent the mean response, with shaded areas equal to the standard deviation. n is the number of cells analyzed. D Representative traces of five individual cells from each treatment and control condition. E Pulse numbers for each individual cell over the duration of the imaging. Bars shown mean ± SD. ***p < 0.001 as assessed by ANOVA. Comparisons between treated and control groups unless otherwise indicated. F Total duration of JNK activity over the 8.5-h experiment. Bars show mean ± SD. ***p < 0.001 as assessed by ANOVA. Comparisons between treated and control groups unless otherwise indicated. G Duration of JNK activity over the first 3 h. Bars show the mean ± SD. ***p < 0.001 as assessed by ANOVA. Ns = not significant. Comparisons between treated and control groups unless otherwise indicated. H–J Western blot images of phosphorylated c-Jun (pJun S73), total c-Jun, and β-actin in response to sustained (H), transient (I), or pulsed (J) anisomycin treatment confirming phosphorylation patterns of c-Jun mirror KTR traces.