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. 2020 Jul 8;10:11239. doi: 10.1038/s41598-020-67724-w

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© The Author(s) 2020

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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ROS staining of neurons for ROS and viability measurements. (a) DIV19 neurons were incubated with MIRB nanoparticles for 2 h (top row) or 24 h (bottom row) at dosages of 0 µg/ml (control), 5 µg/ml, 10 µg/ml or 20 µg/ml. Live neurons were fluorescently observed with a fluorometric intracellular ROS kit (green) particularly sensitive to superoxide and hydroxyl radicals. Scale bar is 100 µm. (b) Plot of the ROS fluorescence intensity per neuron normalized against that of the control. (c) Plot of the number of fluorescent neurons per field of view normalized against that of the control. Each error bar denotes the standard deviation of the mean values of the replicate wells at a given time point and dose.