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. 2024 Dec 28;14:30767. doi: 10.1038/s41598-024-80834-z

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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. 2 — DAC-mediated mtRNA upregulation limits its antiproliferative effects. (A) Glycolytic and mitochondrial ATP content in DMSO or DAC-treated cells was analyzed by Cell Titer Glo. Oligomycin A was used to distinguish the glycolytic ATP from total ATP which was then used to calculate the mitochondrial ATP levels. (B, C) mtDNA copy number (B) or mt-mRNA (C) expression in DMSO or DAC-treated cells analyzed by RT-qPCR. (D) The protein expression of COX1 in DMSO or DAC-treated cells analyzed by western blotting. (E) The expression of POLRMT mRNA in DMSO or DAC-treated cells analyzed by RT-qPCR. (F) Glycolytic and mitochondrial ATP of HCT116 cells treated with DAC measured using Cell Titer Glo. (G) RT-qPCR analysis of mt-mRNA expression in DMSO and DAC-treated HCT116 cells. (H) Western blotting of apoptosis-associated proteins, BCL-2, and NF-κB signaling pathway-associated genes in DMSO and DAC-treated A549 and HCT116 cells. In all experiments, an average of three biological replicates is shown with error bars denoting s.e.m. A Student’s t-test was performed for statistical analysis. * indicates P-value < 0.05, ** indicates P-value < 0.01, *** indicates P-value < 0.001.