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. 2020 Feb 21;27(8):2417–2432. doi: 10.1038/s41418-020-0512-5

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© The Author(s), under exclusive licence to ADMC Associazione Differenziamento e Morte Cellulare 2020

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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Fig. 1 — a Melanoma cell lines respond heterogeneously to single and combination treatment of IZI1551 and Birinapant. Cells were treated for 72 h followed by flow cytometric determination of cell death (propidium iodide positivity). Data shown are means from n = 3 independent experiments. b Synergy scores for treatment combinations, as calculated by Webb’s fractional product method. c Treatment-induced changes in IAP amounts, analysed by Western blotting. Actin served as loading control. Asterisks indicate unspecific bands. Representative results from n = 3 independent experiments are shown. d Apoptotic signalling was studied 24 h after single and combination treatment with IZI1551, Birinapant and Q-VD-OPh (30 µM). Actin served as a loading control. Representative results from n = 3 independent experiments are shown. e Melanoma cell lines die by apoptosis upon combination treatment. Cell lines were treated with 1 nM IZI1551, 1 µM Birinapant, with or without 30 µM Q-VD-OPh. Cells were stained with PI and Annexin V-APC and analysed by flow cytometry. Shown are mean values + SD of three independent experiments.