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. 2021 Jun 3;11:11707. doi: 10.1038/s41598-021-91070-0

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© The Author(s) 2021

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 licence, and indicate if changes were made. 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/4.0/.

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Effect of the combination of LPS with Cu²⁺ and Mg²⁺ on THP-1 cell metabolic activity (a) and gene expression of M1 and M2 markers (b). THP-1 cells were stimulated with a combination of Cu²⁺ and Mg²⁺ and 100 ng/ml LPS for 48 h (strong pro-inflammatory stimulation) or 10 ng/ml LPS for 24 h (mild pro-inflammatory stimulation). (a) Metabolic activity results were represented as percentage relative to an unstimulated control (TCP) and compared with the TCP of each day. Statistical significance was accepted at p < 0.05 and represented with *. (b) M1 markers (TNF-α, IL-1β and CCR7) and M2 markers (IL-10, TGF-β and CD206) were determined by RT-qPCR. Results were normalized against β-actin mRNA and are expressed relative to the mRNA levels of unstimulated THP-1 macrophages. Statistical analysis is in Supplementary Tables S4, S5, S6 and S7.