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. 2019 Mar 18;9:4758. doi: 10.1038/s41598-019-41188-z

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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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Electroporation treatment of three cell lines. HT29 (human colon cancer; top panel), MDA-MB231 (human breast cancer; middle panel) and HDF-n (human dermal fibroblast; bottom panel) treated with calcium electroporation (left), calcium-bleomycin electroporation (middle), and bleomycin electroporation (right). Drug was added at different time points relative to electroporation and experiments were performed at different temperatures (37 °C, 27 °C, and 17 °C). On the HT29 cell line treatment effect of calcium electroporation or calcium-bleomycin electroporation was dependent on temperature and time of drug administration whereas treatment effect of bleomycin electroporation was independent on time of addition but showed dependency on temperature when bleomycin was added before electroporation. Treatment at 27 °C resulted in highest treatment effect for this cell line. Treatment of MDA-MB231 showed neither temperature nor time of addition dependency for any of the treatments investigated. On the HDF-n cell line treatment effect of calcium electroporation or calcium-bleomycin electroporation was dependent on time of drug administration and on temperature whereas treatment effect when treating with bleomycin electroporation was independent on temperature and time of drug administration. Treatment at 37 °C resulted in lowest survival for this cell line. Results from control samples are shown in Supplementary Fig. S1. Mean + SD, n = 5–8, each performed as individual experiments. Obvious outliers have been removed from the data set. Statistical comparisons (two-way ANOVA) are described in the results section.