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. 2017 Nov 2;7:14915. doi: 10.1038/s41598-017-14848-1

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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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SAHA, azacitidine, decitabine, oxaliplatin, fenbendazole and oxibendazole induce HMGB1 release in regular biosensor cells and wild-type cells. U2OS cells stably expressing a GFP-HMGB1 fusion (A–C) or wild-type U2OS cells (D–E) were maintained in control conditions (Ctrl); mitoxantrone (MTX) at 2 µM; suberoylanilide hydroxamic acid (SAHA), fenbendazole (FENB) and oxibendazole (OXB) at 5 (l), 10 (h) µM; azacitidine (AZA) at 15 (l), 30 (h) µM; decitabine (DECI) at 20 (l), 40 (h) µM; and oxaliplatin (OXA) at 50 (l), 100 (h) µM for 24 h or 48 h, followed by assessment of cytoplasmic GFP-HMGB1 fluorescence intensity (A,B) or endogenous HMGB1 level after immunostaining with an anti-HMGB1 antibody and Alexa-fluor 488 conjugated 2^nd antibody and quantification of relative enrichment in the cytosol (D,E). Representative images of biosensor cell line (C) and immunofluorescence (F) are reported (scale bar = 10 µm). Data are shown as means ± SEM (n = 4; *P < 0.05, **P < 0.01, and ***P < 0.001, two-tailed Student’s t test, compared to control cells).