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. 2020 Oct 14;10:17313. doi: 10.1038/s41598-020-73454-w

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© The Author(s) 2020, corrected publication 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 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/.

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Pneumococcal ST615 present 2 Ply pools: cell-wall associated versus cytosolic variant. (A) Determination of haemolytic activity using the standardised sodium deoxycholate-based extraction assay. Serial dilutions were carried out from fresh cultures of bacterial stock at 10⁸ CFU/ml and subsequently incubated with a 4% sheep red blood cell solution. The data are representative of 3 independent experiments (n = 3 replicates, mean ± SEM) (B) Detection and quantification of pneumolysin in the cytosol (left) and cell wall (right) fractions of the indicated pneumococcal strains via Western blot and ELISA analysis using an anti-Ply mouse monoclonal antibody. (C) Determination of the haemolytic activity of the Ply-containing cytosolic fractions (D) Contact-dependent haemolysis was tested in ST615, ST306, TIGR-4, D39 and ply-knockout ST615. PLN-A was used as a negative control.