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. 2018 Oct 2;9:4043. doi: 10.1038/s41467-018-06451-3

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© The Author(s) 2018

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. 4 — NOD2ΔLRR binds RIP2CARD filaments. a Domain organization of NOD2ΔLRR and RIP2CARD constructs used for immuno-gold labelling experiments. Expression of recombinant NOD2ΔLRR was done in sf21 insect cell. b, c Size exclusion chromatography profile (b) and (c) corresponding 12.5% SDS-PAGE showing typical sample obtained from NOD2ΔLRR purification. The size exclusion chromatography profile shows that both tagged (blue) and tag-free NOD2ΔLRR (black) elute in the VV. The dashed rectangular shape indicated the protein fraction used for immuno-gold labelling experiments. d Negative-stain images of VV NOD2ΔLRR, corresponding to the fraction used reconstitution with RIP2CARD, unlabelled (left) and after immuno-gold labelling (right). e, f Example negative-stain micrograph of RIP2CARD filaments with NOD2ΔLRR bound (e) and zoom showing gold-particles (black dots) after immuno-gold labelling against NOD2 (f). Scale bars are 50 nm. VV: size exclusion chromatography void volume