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. 2021 Oct 22;10(11):1088. doi: 10.3390/biology10111088

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Treatment with 6-aminonicotinamide (6-AN) enhances oxidative and endoplasmic reticulum (ER) stress, and affects mitochondrial membrane potential (MMP). (A) Intracellular reactive oxygen species (ROS) levels were determined in both A540 and H460 cancer cells treated with low (10 μM) and high (200 μM) concentrations of 6-AN treatments using flow cytometry. Representative graphs indicate the intensity of DCF fluorescence in 6-AN treated cancer cells normalized to the control. (B) qRT-PCR analysis of mRNA expression of ER stress-related genes such as XBP-1, DDIT3, pERK, and ATF3 following 6-AN treatment in H460 cancer cells under both conditions. (C) Mitochondrial depolarization levels in A549 and H460 cancer cells following low (10 μM) and high (200 μM) concentrations of 6-AN treatments, detected by flow cytometry with the Mitoflow reagent. (D) Representative graphs of levels of Mitoflow intensity in A549 and H460 cancer cells, calculated from panel C as per the indicated treatment conditions. * p < 0.05; ** p < 0.01; *** p < 0.001.