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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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Dependency of induction of bystander inducing potential of tumor cells on the preparation of PAM and on the time of contact between PAM and cells. PAM was generated by treatment of the medium with CAP for 1 min (A) or 3 minutes (B). MKN-45 tumor cells were mixed with PAM (50%) for the indicated times of contact and then washed by three cycles. Increasing percentages of pretreated cells were mixed with untreated tumor cells and the percentages of apoptotic cells in the cocultures were determined 4.5 hours after the beginning of the coculture. The results show that the induction of bystander inducing potential depends on the time of CAP treatment during the preparation of PAM and on the time of contact between PAM and the tumor cells. This is in line with the assumption that CAP generates long-lived species in PAM in a time-dependent fashion and that these compounds need to interact among themselves and with the cells in a time-dependent way.