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. 2017 Sep 14;7:11644. doi: 10.1038/s41598-017-11271-4

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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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Z-stack series of DC migration through epithelia upon fungal infection. (a) Upon fungal infection, DCs (green) sense swollen conidia, germlings (1 h to 6 h, 1^st to 3^rd panel) and hyphae (24 h) (red) at the apical side of differentiated epithelia (white) and transmigrate over time to the apical side and upon destruction of the epithelium. Actin is illustrated in white, conidia/hyphae in red, nuclei in blue and DCs in green. (b) The image shows an x-y projection with the respective side views of DCs (green) interacting with hyphal filaments (red) at the surface of destroyed epithelial cells (white) at the 24 h time point. Bars, 10 µm. CLSM was performed capturing at least 3 different areas each containing more >25 cells per condition. (c) Percentages of apically visible DCs over time (1 h to 24 h) in infected DC/NHBE co-cultures are depicted. Significantly more DCs were found at 3 h, 6 h and 24 h at the apical layer compared to background percentages of control cells. Control cells remained at the basolateral side and no increase in DC numbers at the apical layer was detected. Numbers of DCs counted were ~125. Significant differences between infected and control co-cultures were calculated using an unpaired Student’s T test and the Graph Pad Prism software.