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. 2018 May 22;8:7985. doi: 10.1038/s41598-018-25865-z

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© The Author(s) 2018

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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TatD is associated with pneumococcal extracellular vesicle fraction. (a) The extracellular vesicle (EV) fraction from S. pneumoniae strain D39 was immunoblotted with mouse anti-rTatD polyclonal antibody. We probed for pneumolysin as a positive control as pneumolysin (53 kDa) has previously been demonstrated to be present in the EV fraction. This experiment was done three times. Data from a representative experiment is shown. (b) DNase assay was performed with EVs isolated from D39 and D39ΔtatD strains as described in the Material and Methods. Bovine pancreatic DNase I (BP DNase) and PBS served as the positive and negative control, respectively. This experiment was performed thrice and data from a representative experiment is presented. The full-length blot (panel a) and gel (panel b) can be viewed in Supplementary Fig. S7. (c) The DNase activity present in the EV fraction (1 μg) from D39 and D39ΔtatD strains was quantitated using picogreen assay. The experiment was performed twice. The data from a representative experiment is shown. Two tailed unpaired t test was performed and error bars represent mean ± sem of duplicates.