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. 2018 Jan 9;8:144. doi: 10.1038/s41598-017-18644-9

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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CANE sensitizes A549 cells to follow ER stress leads to mitochondria-mediated apoptotic pathway. (a) Western blotting of ER stress markers, β-actin used as loading control. Docetaxel (20 μg/ml) was used as a positive control while the surfactant-polysorbate 80 mix devoid of carvacrol was also used as a control. Densitometry analysis of IRE1-α, PERK, and ATF-6 was determined by Image J software. (b) Evaluation of intracellular Ca⁺² accumulation. (c) Mitochondrial membrane potential of A549 cells treated with CANE (25-100 μg/ml). Fluorescent images were captured at 20X magnification [scale bar = 0.1 mm]. Image J software was used to determine the mean fluorescence intensity. (d) Effect of CANE on expression of Bax, Bcl2, and Cyt C in A549 cells. (e) Cyt C level in CANE-treated cells measured by spectrophotometric method. (f) Effect of CANE on expression of cleaved caspase-9, and 3 in A549 cells. Each value in the graph represents as the mean ± SD of three independent experiments. Values with different superscripts differ significantly from each other (p < 0.05).