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. 2022 Aug 19;6(8):979–991. doi: 10.1038/s41551-022-00922-1

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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 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, Rationale and assay schematic. Created with BioRender.com. b,c, Western blot densitometry of LAM (b) and LprG (c) expression in cytosolic (Cyto), EV and cell membrane (CM) factions of macrophage cultures infected with the indicated Mtb strains. Data indicate mean ± s.d., n = 3 biologically independent repeats; P values obtained by one-way ANOVA with Dunnett’s post-test. a.u., arbitrary units. d, Western blot analysis of LAM and LprG from CFP samples of patients with sequence-confirmed M. smegmatis, Mtb, M. avium, M. intracellulare and M. kansasii infections. e,f, EV-ELISA for LAM on EVs from Mtb-infected macrophages (e) and BMVs isolated from Mtb culture medium (f) by ultracentrifugation when captured by host EV-specific antibody. g, EV-ELISA for LAM and LprG on serum EVs of non-human primates with pulmonary TB (PTB), LTBI or their healthy controls (Ctrl) (mean ± s.d. of three technical replicates). h, EV-ELISA for LAM, LprG and integrated LAM and LprG (LAM+LprG) expression on isolated EVs from serum of children with TB (N = 10) and without evidence of TB (Ctrl; N = 5). Mean ± s.e.m., P values obtained by two-sided Mann-Whitney U test.