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. 2024 Oct 30;14:26051. doi: 10.1038/s41598-024-77692-0

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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. 3 — Effect of mitochondrial inhibitors and C-terminus mutations on CRY stability. (A) Left Panel: prolonged (2 h) light exposure effectively degrades CRY proteins in S2 cells treated with vehicle (DMSO, EtOH) or rotenone (Rot), whereas complex III (Ant, Myx) and V (Oli) inhibitors block light-dependent CRY degradation. Right panel: compared to wildtype (wt) CRY, levels of C523S, E530Q, F534L and F534W mutant proteins are reduced, and light exposure further destabilizes C523S, F534L and F534W proteins. (B) Relative protein levels were quantified from more than 3 Western blots. Asterisk denotes significant difference between dark and light, by Student’s t-test, p < 0.05. (C) Complex III and V inhibitors do not block degradation of CRY^m (m) proteins. Error bar denotes standard error of the mean, n > 3. Different letters above the bar denotes significant difference (one-way ANOVA, post hoc Tukey test, p < 0.05). Full-length blots are presented in Supplemental Figure S11.