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. 2022 Mar 10;12:3906. doi: 10.1038/s41598-022-07804-1

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© The Author(s) 2022

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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The NOD2 p.E54K variant leads to abrogation of both PGN-dependent and -independent signaling. (A) NF-kB luciferase reporter activity upon challenge with L18-MDP (7 h) in HEK293T cells overexpressing NOD2 wild-type (WT) or indicated mutants. (B and C) Quantitative RT–PCR analysis and ELISA of IL8 expression upon stimulation with L18-MDP (B) or tunicamycin (C) in heterologous HCT116 cells. (D) Representative immunoprecipitation of FLAG-tagged NOD2 (n = 3) on HEK293T cells that were transiently transfected with Flag-NOD2 WT or indicated mutants alone or along with WT RIPK2. (E) Representative TUBE assay (n = 2) from L18-MDP-treated heterologous HCT116 cells. Data represent mean ± SEM of three (A and B) or five (C) independent experiments. P values for each treatment group are calculated in comparison to WT. WCL, whole cell lysate; IP, Immunoprecipitates.