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. 2022 Mar 9;7:71. doi: 10.1038/s41392-022-00888-1

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© The Author(s) 2022, corrected publication 2023

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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Fig. 3 — Effect of SHK on NEMO/IKKβ/NF-κB signaling. a The protein levels of NEMO, p-IKKβ/IKKβ, and p-IκBα/IκBα in LoVo cells treated with the indicated doses of SHK were analyzed by western blotting. b The protein levels of NEMO, p-IKKβ/IKKβ, p-IκBα/IκBα protein in tumor tissues were analyzed by western blotting. c LoVo cells were co-transfected with pNFκB-luc containing a Renilla luciferase reporter (as internal control) for 24 h and then treated with SHK for 48 h. Luciferase activity was determined using a dual-luciferase reporter assay system. d The IKKβ kinase activity was assessed using a spectrophotometric quantitative detection kit in LoVo cells. The relative activity was calculated using the provided formula. e The subcellular localizations of p50/p65 in LoVo cells following 48 h of treatment with SHK was examined by confocal microscopy analysis. The protein levels of p65 and p50 in nucleus (f) and whole-cell lysates (g) were measured by western blot analysis. Lamin B1 and β-actin were used as controls for sample loading. h The binding of p65 and p50 to a COX-2 promoter probe was analyzed using a streptavidin-agarose pull-down assay. The data are presented as the means ± SD of at least three separate experiments. (*p < 0.05, **p < 0.01, SHK treatment group vs. vehicle control group)