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. 2020 Mar 31;10:5718. doi: 10.1038/s41598-020-62749-7

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© The Author(s) 2020

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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C. longa and A. hookeri co-treatment inhibits NF-κB and COX-2 expression. (A) NF-κB is downregulated in the nucleus and COX-2 in the cytoplasm. NF-κB and COX-2 are not expressed in the control group and only the nucleus is stained with DAPI (blue spots). Carrageenan stimulates the expression of NF-κB in the nucleus and that of COX-2 in the cytoplasm. MSM suppresses the expression of NF-κB and COX-2 induced by carrageenan, but the expression of NF-κB (arrow) and COX-2 (arrowhead) observed is minimal. C. longa and A. hookeri co-treatment significantly inhibits the expression of NF-κB and COX-2. (B) The graph and scores present the immunofluorescent results using the image analyzing program in the K1-Fluo confocal microscope. MSM and C. longa and A. hookeri co-treatment statistically significantly control NF-κB and COX-2 expression effectively. (C) Compared to the density of NF-κB in the nucleus and cytoplasm, the pattern is similar to that observed in the immunofluorescence assay. C. longa and A. hookeri co-treatment blocks the translocation of NF-κB into the nucleus from the cytoplasm and inhibits the expression of COX-2 in the cytoplasm. (D) Depending on the image analysis of the western blotting of NF-κB (in nucleus and in cytoplasm) and COX-2 (in the skin tissue), the density graph based on each band per β-actin was presented using the Image J program. (E) MSM and C. longa and A. hookeri co-treatment significantly inhibited carrageenan-induced iNOS in the serum. The downregulation effect of 500 mg/kg C. longa and A. hookeri co-treatment is similar to that of the control. All values are presented as mean ± standard deviation. Arrow, NF-κB; arrowhead, COX-2. Magnification, 1000×. Scare bar, 50 μm. CON vs. ^*p < 0.05; CON vs. ^**p < 0.001 Carrageenan vs. ^$p < 0.05; Carrageenan vs. ^$$p < 0.001; MSM vs. ^#p < 0.05. Table in Fig. 4. The ratio of NF-κB p65 vs. nucleus or COX-2 vs. nucleus. All values are presented as mean ± standard deviation. CON vs. ^*p < 0.05; CON vs. ^**p < 0.001; Carrageenan vs. ^$p < 0.05; Carrageenan vs. ^$$p < 0.001; MSM vs. ^#p < 0.05.