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. 2020 Feb 3;10:1653. doi: 10.1038/s41598-020-58569-4

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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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Spheroid response to RTHT. (A,C) Growth curves for HCT116-4000 and CAL27 spheroids following RTHT treatment 96 h after seeding. Combinations of HT (16 to 560 CEM₄₃) with 2 Gy (A) or 5 Gy (C) are shown. (B,D) Mean PI staining intensity as a function of time after RTHT exposure (HCT116-4000 spheroids). (E) Phase contrast images overlaid with PI fluorescence snap-shots from time-lapse imaging of representative HCT116-4000 spheroids treated with 5 Gy + 80 CEM₄₃ or 2 Gy + 40 CEM₄₃. Means and standard errors of the mean of at least three independent repeat experiments are shown in A to D ( $n \geq 3$ ).