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. 2023 Mar 25;13:4889. doi: 10.1038/s41598-023-31990-1

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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 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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Experimental modulation of cell cycle progression affects the background autofluorescence. HCT 116 cells were synchronized to G0/G1 phase by cultivation to the full confluency (red line plots) or arrested in the G2/M phase with nocodazole treatment (blue line plots). Control HCT 116 cells were cultivated in standard subconfluent conditions (green line plots; see Methods for details). Dead cells were excluded from analysis using LIVE/DEAD Fixable Dead Cell Stain Kit. The cell cycle was then analyzed in both native (Vybrant DyeCycle Violet) and fixed (FxCycle Far Red Stain) conditions. Together with background fluorescence. The numbers of G0/G1 and G2/M phases represent a percentage of cells. The values of G0/G1 and G2/M phase represent the proportion of cells. Data are representative from two independent repetitions. MFI median fluorescence intensity.