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. 2020 Apr 6;10:5969. doi: 10.1038/s41598-020-62846-7

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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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Streptococcus pneumoniae D39 aggregation on HMEECs monolayer detected by PNA probe and fluorescence in situ hybridization (FISH). (a) Confocal microscopic images of HMEECs untreated, cell nuclei stained with Hoechst dye were visible blue. (b) Confocal microscopic image of HMEECs treated with S. pneumoniae only, bacteria were labelled with PNA probe (green) on HMEECs and on the bottom surface. (c) Confocal microscopic image of HMEECs treated with UP + S. pneumoniae (co-treatment), bacteria formed aggregations (green) on HMEECs and were labeled with PNA probe. Images are 20×.