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. 2017 Oct 12;7:13065. doi: 10.1038/s41598-017-13588-6

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© The Author(s) 2017

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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Biofilms formed by C. glabrata elicit NET release. (a) The biofilm-forming capacity of C. glabrata isolates was estimated by XTT assay after 24 h of growth. Assays were performed in triplicate on 3 days and representative data is shown with SD. (b) C. glabrata biofilms (24 h) were co-cultured with human neutrophils and NET release was estimated by Sytox Green detection of free DNA. Results were normalized to the positive control, PMA, and data from 5 experiments performed in triplicate were combined. Neutrophil responses to Candida were analyzed by ANOVA with pairwise comparison to the untreated neutrophil control, *P < 0.05, SEM shown. (c) Neutrophil interactions with C. glabrata at 4 h were imaged with scanning electron microscopy. Measurement bars represent 10 µm and 1 µm for 2,000x and 10,000x images, respectively. (d) The neutrophil response to C. glabrata biofilms was imaged with scanning electron microscopy at various time points over 4 h. Measurement bars represent 1 µm for 10,000x images.