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. 2019 Mar 13;9:4376. doi: 10.1038/s41598-019-40771-8

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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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Changes in the expression levels of selected proteinous virulence factors in C. albicans during contact with P. gingivalis. After a 3 h contact between C. albicans and P. gingivalis cells in the simultaneous model of interaction under anoxic and normoxic conditions, total RNA was extracted from fungal cells using TRI reagent treatment. Real time quantitative PCR was then performed with a SYBR green detection assay. The selected genes encoding fungal surface protein Hwp1, adhesins of the agglutinin-like family, Als3 and Als7, aspartic proteases, Sap3, Sap6 and Sap9, and a cytosolic/‘moonlighting’ protein enolase (Eno1) were analyzed. The C_T values (the average threshold cycle) were normalized to that of the ATC1 gene. To quantify gene expression, comparative C_T was used and the relative expression was determined using the 2^−ΔΔCT method⁷¹. Results are representative of three independent experiments and are expressed as means ± SD (standard deviation); n = 3. *P < 0.05; **P < 0.01; ***P < 0.001. The analysis was performed using GraphPad Prism software (GraphPad, LaJolla, CA, USA) with an one-way ANOVA test.