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. 2021 Mar 16;11:6008. doi: 10.1038/s41598-021-85387-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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The confirmation of measuring chelatable labile iron in vitro. HepG2 cells were loaded with 200 μM of iron using FAC. Cells were lysed and these lysates were treated with either deferoxamine (DFO), deferiprone (DFP) and deferasirox (DFX) at 50 μM overnight. (A) LIP—measured using 5 mM ferene and 10 mM ascorbic acid in lysates. Chelatable labile iron were measured in cell lysates with iron chelator treatments. Labile iron was measured in cell lysates without iron chelator treatments. (B) Total iron—measured in nitric acid digested cell lysates followed by the ferene-based assay for total iron (5 mM ferene and 1 M ascorbic acid). One-way ANOVA was performed with Tukey’s correction using GraphPad Prism.