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. 2019 Oct 25;9:15336. doi: 10.1038/s41598-019-51703-x

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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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The length of S/G2/M is affected by let-7 overexpression and knockdown. Real-time assessment of cell cycle phase length by live-imaging using the fluorescent reporter FUCCI. (A) Schematic of the FUCCI reporter. Cells express Cdt-1 RFP (red) in G1. Upon entry into S-phase, cells begin to express Gem-GFP and are yellow for a short period before Cdt-1 RFP is degraded and the cells are green. (B) Representative example of a movie assessing the length of S/G2/M in control HER10-FUCCI cells. The cell being tracked (red arrow) is red at 0 h. After 2.5 h, Gem-GFP becomes detectable (green arrows). The cell remains green for 17.1 h before dividing into two daughter cells (white arrows; see Movie 3). (C) Representative example of a movie assessing the length of S/G2/M in let-7 Ant-transfected HER10-FUCCI cells. In this example, the Gem-GFP+ phase is only 10.7 h (green arrows) before the cell divides into two daughter cells (white arrows; see Movie 4). (D) Representative example of a movie assessing the length of S/G2/M in let-7 mimic transfected HER10-FUCCI cells. In this example, the Gem-GFP+ phase is extended to 23.3 h before the cell divides (white arrows; see Movie 5). (E) Quantification of all HER10-FUCCI live imaging. (n = 3 movies per condition, with approximately 30 cells tracked per phase (G1 or S/G2/M), per condition).