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. 2020 Jun 15;10:9640. doi: 10.1038/s41598-020-66531-7

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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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Production of the antigen. The pictures show western blots of cell-free extracts ofAg85B-ESAT6-DC(AgE6-DC) expressing strains harvested 3 hours after induction. Sample sizes were adjusted to the OD₆₀₀ of the harvested culture, meaning that all samples represent approximately equal amounts of cells. Lanes: M, molecular mass markers (masses are indicated in kDa); pEV, strain harboring empty vector; cyt, strain harboring vector for intracellular localization (expected mass of the fusion protein is 41 kDa); lipo (48 kDa), cwa2 (69 kDa) and LysM (66 kDa), cell-free extracts of strains harboring various plasmids for anchoring (expected masses between parenthesis). The data presented are from one representative experiment, out of at least three experiments in total. Parts of the lanes marked “lipo” have been published previously³⁰, except in the case of L. sakei.