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. 2022 Dec 5;20:270. doi: 10.1186/s12915-022-01472-3

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Proposed EV biogenesis mechanisms in gram-positive bacteria. A Proposed models for EV biogenesis in gram-positive bacteria. B, C Representative (B) multi-color STORM and (C) TEM images of S. aureus with EVs for each mechanism. Red: WGA. Green: Nile red. Yellow arrow: expanded IWZ. D Comparison of the cell wall density for each mechanism observed from STORM images. Averaged localization number in the cell wall was measured as the cell wall density from STORM images. E Relationship between the contrast and the curvature of the cell wall measured from TEM images. The fitting line obtained by the exponential function shows an inverse relation. F Population ratio of S. aureus producing EVs by each mechanism observed from multi-color STORM and TEM images (mean±SD; n=~1000 from STORM images, n=~80 from TEM images). G Comparison of the diameter of EVs produced by each mechanism observed from STORM and TEM images (mean±SD; n=~30). White arrows: observed EVs. Scale bars: 1 μm in B and 200 nm in C