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. 2019 Mar 13;9:4346. doi: 10.1038/s41598-019-39554-y

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© The Author(s) 2019

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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Effects of CM14 on the virulence-related phenotypes of V. vulnificus. (a,b) The V. vulnificus strains grown along with CM14 as indicated or DMSO (control) were harvested and fractionated for further analyses. (a) The transcript levels of vvhA, rtxA, and plpA in the total RNA of the cells were quantified by qRT-PCR and expressed using each transcript level of the wild type in the presence of DMSO as 1. (b) Hemolytic activities of the culture supernatants were determined against human erythrocytes and expressed using complete hemolysis by 5% Triton X-100 as 100%. (c) Morphological changes of HeLa cells infected with the V. vulnificus strains along with CM14 (50 μM) or DMSO (control) were photographed. Scale bars, 100 μm. (d) Cytotoxicity was determined using LDH activities released from INT-407 cells infected with the V. vulnificus strains along with CM14 as indicated and expressed using the LDH activity from the cells completely lysed by 5% Triton X-100 as 100%. Error bars represent the SD from three independent experiments (a,b) and from the representative of three independent experiments (d). Statistical significance was determined by the Student’s t-test (a) and by one-way ANOVA (b,d) (***p < 0.0005; **p < 0.005; *p < 0.05; ns, not significant). WT, wild type; hlyU, hlyU mutant.