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. 2023 Jul 11;14(1):2228657. doi: 10.1080/21505594.2023.2228657

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© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 5. — Sau-41 represses S. aureus haemolytic activity. (a) Schematic illustration of the mutation of the PSMα4 start codon. (b) Hemolytic ring of the RN4220 strain carrying Sau-41/Sau-41^mt overexpression, knockdown or empty vector plasmid grown on sheep blood agar overnight. (c) Hemolytic activity of the RN4220 strain carrying Sau-41/Sau-41^mt overexpression, knockdown or empty vector plasmid determined by incubating bacterial supernatants with 3% human red blood cell. PBS was used as a negative control, and H₂O was used as a positive control. (n = 3/group) (d) Transcript levels of hla, hld, psmα, psmβ, hlgA, hlgB, and hlgC were investigated by qPCR in strain RN4220 carrying pRMC2 or pRmc2_sau-41 (n = 3/group). (e) Volcano plot showing the haemolytic factor transcriptional differences between strain RN4220 carrying pRMC2 or pRmc2_sau-41 and their corresponding adjusted p value (q value). (f) the transcriptional activity of hla in strain RN4220 was assessed by analysing cells that harboured plasmids capable of either overexpressing or knocking down Sau-41 (n = 3/group). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Sau-41⁺: RN4220 carrying the pRmc2_sau-41 plasmid, Sau-41^mt+: RN4220 carrying the pRmc2_sau-41mt plasmid, Sau-41^kd: RN4220 carrying pSd1_sau-41, wild-type: RN4220 carrying the pRMC2 empty vector.