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. 2019 Sep 26;9:13931. doi: 10.1038/s41598-019-50329-3

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© The Author(s) 2019

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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Basic scheme of the bystander experiments performed in this study (Figs 14–19). Tumor cells were treated with CAP for 1 min and then either further cultivated in the same medium for additional 25 min, and then washed (A,B), or washed immediately after CAP treatment (C,D). Pretreated cells from assays A and C were added at increasing percentages to untreated tumor cells immediately after the washing step. Pretreated cells from assays B and D were subjected to 25 min incubation after the first washing step, were then washed and added to untreated tumor cells at increasing percentages. As indicated in this figure and specified in the respective legends, defined inhibitors or scavengers could be applied at various steps. The titration of the pretreated cells, in combination with their potential for bystander signaling, allows to quantitatively determine the number of cells that had obtained an “imprint” during a specific, experimentally defined step. This allows to conclude back on the dynamics of the underlying processes. The use of inhibitors and scavengers thereby allows to define the chemical biology involved in these dynamic interactions.