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. 2018 Sep 20;8:14144. doi: 10.1038/s41598-018-32603-y

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© The Author(s) 2018

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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P. gingivalis invasion of human oral keratinocytes. CTV-labeled TIGKs were infected with various CFSE-labeled strains of P. gingivalis (MOI = 200) for 90 min. Metronidazole was added for an additional 1 h, following which the cells were fixed and analyzed by flow cytometry. Cells were first gated on the basis of a forward scatter/side scatter (FSC-A/SSC-A) plot (a). The events were then visualized using FSC-A/FSC-H dot plot, and single cells were gated (b). TIGKs were identified on the basis of CTV positivity (c). TIGKs invaded by P. gingivalis were subsequently defined as the CFSE⁺ cells within the CTV⁺ keratinocyte population (d). Overall, 20,000 events were analyzed, and the results are presented as the percentage of TIGKs infected with P. gingivalis wild-type (e), Δppad (f), and C351A (g).