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. 2021 Dec 1;12:6998. doi: 10.1038/s41467-021-27388-0

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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 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. 1 — a Schematic representation of the Photorhabdus laumondii T6SS gene cluster. Genes encoding T6SS core components of T6SS are identified by their Tss corresponding letter (e.g., A for TssA). A close-up emphasizing the vgrG-eagR-rhs1-imm1 genetic locus is shown below (vgrG, orange; eagR, blue; rhs1, red; tri23, green). b and c Pull-down assays. Total cell extracts (T) from E. coli BL21(DE3) cells producing various combinations of His₆-tagged VgrG (^HVgrG, V), Strep-tagged EagR (^STEagR, E), and FLAG-tagged Rhs1 (Rhs1^FL, R) were subjected to purification on streptactin-agarose beads. Strep-tagged and co-precipitated proteins were eluted with desthiobiotin (IP). Total and IP fractions were analyzed by SDS-PAGE and co-purified proteins were stained by Coomassie blue (b) or immunodetected using anti-His, anti-Strep, and anti-FLAG antibodies (c). Molecular weight markers (Mw, in kDa) are indicated on the left. Pull-down assays have been performed at least in triplicate, and a representative experiment is shown. d Schematic representation of the VgrG (orange), EagR (blue), and Rhs1 (red) interaction network based on pull-down experiments.