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. 2019 Nov 12;9:16564. doi: 10.1038/s41598-019-52975-z

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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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S. aureus internalization is increased within A549 epithelial pulmonary cells in the presence of P. aeruginosa. (A) Adhesion of S. aureus onto epithelial cells. A549 cells were infected at MOI 10:1 for S. aureus monoculture and 20:1 for S. aureus/P. aeruginosa coculture. After 2 hours of contact, cells were washed with phosphate buffer saline (PBS) to remove unattached bacteria and lysed with sterile water. Supernatants were plated on MSA to count S. aureus. The results are represented as the percentage of inoculum that adhered. (B) Internalization of S. aureus within epithelial cells. After 2 hours of contact, cells were treated with antibiotics and lysostaphine for one hour, lysed with sterile water and bacteria plated on MSA. The results are represented as the percentage of adhered cells that have internalized. All values represent the mean + standard deviation from three independent experiments with three strain pairs (SA27-PA27, SA31-PA31 and SA69-PA69). Statistical significance was determined by unpaired t-test (**P < 0.01).