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. 2020 Feb 26;10:3495. doi: 10.1038/s41598-020-60449-w

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© The Author(s) 2020

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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Invasion of HOK-16B cells and induction of apoptosis by oral E. coli strains. HOK-16B cells were infected with oral E. coli strains and S. salivarius (Ss) at an MOI of 1000. As a positive control (PC), cells were treated with 0.5 µg/ml staurosporine. (A,B) Cell viability and proliferation were measured by trypan blue enumeration. *p < 0.05 compared to untreated cells by t-test. (C) The apoptotic stage was measured by flow cytometry using Annexin V-PI staining. *p < 0.05 compared to untreated cells by t-test. (D) HOK-16B cells were infected with bacteria at an MOI of 1000 for 2 hours, and the infected cells were treated with gentamicin for 1 hour. The cells were then lysed immediately or further incubated for 24 hours in antibiotic-free medium. The cell lysates were cultured on LB agar plates. *p < 0.05 compared to Ss by Mann-Whitney U test, ^#p < 0.05 by 2-tailed Wilcoxon signed-rank test. All graphs present the mean and standard deviation of two experiments performed in triplicate.