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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. 4 — Effect of mitochondrial inhibitors on the transcriptional repressor activity of CRY. (A) In HEK 293 T cells, CLK-CYC activates per-luc2p transcription, which is partially repressed by Drosophila CRY or PER. Rotenone (Rot) and antimycin (Ant) have no effect on per-luciferase activity. ns denotes no significant difference by one-way ANOVA. (B) In HEK 293 T cells, CRY mutants are less effective than wildtype CRY in repressing per-luc2p reporter activity, most likely due to their lower expression levels. Error bar denotes standard error of mean, n = 3. Asterisk denotes significant difference (between wt and each mutant group) by Student’s t-test, p < 0.05. (C) In S2 cells, CLOCK-BMAL1 activates per1-dluc transcription, which is repressed by wildtype mCRY1 (wt). The mCRY1 D387A mutation (equivalent of Drosophila CRY^b) abrogates such repression. Mitochondrial inhibitors rotenone (Rot) and antimycin (Ant) have no effect. ns denotes no significant difference by one-way ANOVA.