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. 2019 Dec 16;9:19162. doi: 10.1038/s41598-019-55631-8

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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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Heme-induced apoptosis of iPSCs is diminished by NRG-1. iPSCs were treated with heme for 8 hours, and NRG-1 was then added for 18 hours. Cell viability was determined using the CCK-8 assay (A), while the apoptotic/necrotic cell number was assessed using flow cytometry analysis (B,C). (A) Cell viability decreased significantly when iPSCs were treated with heme. NRG-1 reduced the effects of heme. (B) The number of apoptotic cells increased with heme compared to basal conditions. The addition of NRG-1 to heme treatment decreased not only the number of apoptotic but also the number of necrotic cells. CPT was used as a positive control for apoptosis. The data shown are the mean values + standard error (SE) (n = 3 experiments) (a: p < 0.05 compared to basal control; b: p < 0.05 compared to heme). (C) Representative Annexin V/7-AAD flow cytometry results for cells undergoing apoptosis and necrosis under heme treatment. The percentage of cells in each quadrant is shown. Taken together, these results show that heme induced iPSC injury, while the addition of NRG-1 reduced this effect by reducing the apoptotic/necrotic cell number, thus increasing the number of live cells.