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. 2018 Feb 6;8:2494. doi: 10.1038/s41598-018-20861-9

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© The Author(s) 2018

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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Cell survival with a long-term, toxin-induced inhibition of protein synthesis. Vero-d2EGFP cells were incubated for 36 h with (a) 0.05 ng/mL of ricin, (b) 0.01 ng/mL of Stx1, (c) 1.0 ng/mL of EtxA, or (d) 0.05 ng/mL of Dtx. Left column: Representative images were taken at 200 × magnification. Cell viability, as assessed by MTS assay (n = 3, avg. ± std. dev.), is indicated. Center column: Red and blue lines (corresponding to the color-coded toxin concentrations in Supplementary Figure S1) were generated from cytofluorometry analysis of the adherent subpopulation of toxin-treated Vero-d2EGFP cells. Unintoxicated parental Vero cells (black lines) and unintoxicated Vero-d2EGFP cells (green lines) were also processed for each condition. Right column: Cell viability was recorded by cytofluorometry analysis of annexin V and 7-AAD staining (blue), while EGFP fluorescence was recorded by cytofluorometry (grey) or with a plate reader (black). Results are presented as percentages of the values obtained from unintoxicated cells. All measurements in the matched center and right columns were performed on the same population of cells.