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. 2019 Oct 7;9:14362. doi: 10.1038/s41598-019-50692-1

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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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The effect of purinergic antagonists on calcium influx induced by Shiga toxin in HeLa cells and human platelets. (A) Calcium influx was measured in HeLa cells preincubated with NF449, suramin or phosphate buffered saline (PBS) vehicle, stimulated with Shiga toxin 1 (Stx1) or Hank’s balanced salt solution (HBSS) (groups differentiated by icon colors) and imaged by fluorescence microscopy. Results are presented as mean fluorescent change of all cells in the field of view (median and range). The color of the asterisks corresponds to the color of the icon in comparison to Stx1. The absence of asterisks indicates that statistics was not significant. (B-C) Human platelets (n = 3 donors) were preincubated with NF449 or PBS vehicle followed by Stx1 (B) or Stx2 (C) and O157LPS (to enable platelet activation by Shiga toxin) or PBS vehicle. Data is presented as the initial fluorescence subtracted from fluorescence after 2 minutes and the bar denotes the median fluorescence. RFU: relative fluorescent units, ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, two-way repeated measure ANOVA (panel A) and Kruskal-Wallis test (panels B and C).