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. 2023 May 5;14(5):306. doi: 10.1038/s41419-023-05806-z

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© The Author(s) 2023

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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Fig. 2 — a Representative live cell confocal fluorescence microscopy images showing the spheroid morphology of HCT116 and SW620 after 3 days of 5-FU treatment. Cells were stained with either cell tracker CMFDA (green) or CMRA (red) fluorescent probes. Scale bars = 100 μm. b–e 5-FU response of CMS1 and CMS4 cells in spheroids. Monocultured (white bars) and cocultured spheroids (gray bars) of b HCT116 and SW620, c HCT116 and MDST8, d LoVo and SW620 and e LoVo and MDST8 were exposed to different concentrations of 5-FU for 3 days. Cell viability was measured by means of image analysis quantifying the fluorescence intensity of cell trackers that represent the cell number, and normalized to the vehicle control. The bars represent the average of viability and the error bars represent the standard deviation (n = 3). Statistical significance was calculated using a one-way ANOVA followed by Student’s t-test. P-values of <0.05 and 0.01 were considered significant (*) and highly significant (**), respectively, when compared to the monoculture.