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. 2019 Mar 19;9:4863. doi: 10.1038/s41598-019-41273-3

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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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(a) Fluorescence images of early- (top panel) and late-stage (bottom panel) HepG2 spheroids with necrotic edges (red) after 24 hours of sorafenib exposure at a concentration of 0 µM and 100 µM. Scale bar, 200 µm. (b) Fluorescence intensity profile of early- and late-stage HepG2 spheroid images after treatment of 100 µM sorafenib for 24 hours. (c) Drug diffusion distance of sorafenib at a concentration of 100 µM after 24 hours exposure with respect to spheroidal age of HepG2 spheroids. Error bars represent ± SD (n = 4). (d) Inward diffusion of the fluorophore rhodamine B of early and late-stage HepG2 spheroids. (e) Fluorescence intensity fold change of HIF-1α immunohistochemical stained HepG2 spheroids relative to day 3 after cultivation time of 6, 12 and 18 days post-seeding. Error bars represent ± SD (n = 4) and *p < 0.05.