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. 2015 Sep 22;17(1):130. doi: 10.1186/s13058-015-0640-x

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© Mehmeti et al. 2015

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Pathogen-associated molecular patterns (PAMPs) and damage-associated molecular pattern (DAMPs) induce cytokine expression in human breast cancer cells in vitro. a Relative expression of IL-6 and IL-8 mRNAs after 6 h stimulation with lipopolysaccharide 1 (LPS1) and LPS2, using quantitative real-time PCR (QPCR) on mRNA from the cell lines indicated; n = 6. Error bars standard error of the mean (SEM); ***P <0.001 (analysis of variance (ANOVA)). b Release of IL-6 and IL-8 after stimulation with LPS1 and cycloheximide for 6 h, using ELISA on supernatants from stimulated MDA-MB-231 cells; n = 3. c-e Relative release of IL-6 and IL-8 after stimulation with LPS1, LPS2, the DAMP HMGB1 or IL-1β for 6 h, using ELISA on supernatants from stimulated MDA-MB-231 cells (c), SUM-149 cells (d) and SUM-159 cells (e); n = 10. Error bars SEM; *P <0.05, ***P <0.001 (ANOVA). f Dual luciferase reporter assays of MDA-MB-231 cells transfected with an NFκB reporter. TK-Renilla was co-transfected as control (Ctrl). LPS1, LPS2, HMGB1 or IL-1β was added to stimulate NFκB activity as described in “Methods”; n = 14. Error bars SEM; ***P <0.001 (ANOVA). g The relative release of IL-6 and IL-8 after stimulation with the DAMP S100A9 for 20 h, using ELISA on supernatants from stimulated MDA-MB-231, SUM-149 and SUM-159 cells; n = 6. Error bars SEM. *P <0.05, ***P <0.001 (ANOVA). h Transient transfection of pDUO-MD2/hTLR4 for 72 h in MCF-7 cells induces release of both IL-6 and IL-8 measured by ELISA; n = 6. Error bars SEM; *P <0.05, ***P <0.001 (Students t test)