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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 treatment affects ERBB4 and NRG-1 expression in iPSCs. iPSCs were treated with heme for 18 hours. The number of cells positive for ERBB4 was assessed using flow cytometry analysis. The ERBB4 and endogenous NRG-1 (eNRG-1) protein expression levels were determined using Western blotting. (A) Representative flow cytometry results illustrating the percent of iPSCs that express ERBB4. (B) The number of cells positive for ERBB4 increased as the heme-induced injury level increased. (C) Cell lysates from iPSCs treated with various heme doses were immunoblotted for ERBB4 and NRG-1. Western blot densitometry analysis was performed using the ImageJ program (full blots are shown in the Supplementary Information file, Supplementary Fig. S1). The results show that NRG-1 expression levels increased as the heme concentration increased, while ERBB4 expression decreased in a similar manner (a: p < 0.05 compared to basal conditions; b: p < 0.05 compared to 10 µM treatment).