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. 2017 Nov 22;7:16011. doi: 10.1038/s41598-017-16197-5

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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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Co-staining of DENV and HMGB1 in the liver. Liver sections from dengue fatal case (case 4) and a non-dengue case were processed for in situ hybridization and IHC assays. DENV was detected by a probe that anneals to a conserved sequence within the viral RNA negative strand and HMGB1 was assessed by immunohistochemistry assay. In the dengue case, the figure shows a diffuse area of hepatic steatosis with presence of DENV RNA in Kupffer cells (KC) and in sinusoidal borders of hepatocytes. Expression of HMGB1, when considering the dengue case, occurred within the nucleus as well as the cytoplasmic regions. DENV RNA and HMGB1 signals co-stained within the cytoplasm of hepatocytes and Kupffer cells. In the control case, KCs were negative for DENV and, in general, the HMGB1 expression was marginal and predominantly in nuclear regions.