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. 2023 May 13;12(10):1380. doi: 10.3390/cells12101380

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Nup62-Scarlet disassembly from NPCs precedes centrosome splitting at the onset of mitosis. Selected time points from a live-cell imaging (Video S8) show a cell co-expressing the Nup62-Scarlet (magenta) and CP75-GFP (green) knock-in construct. CP75-GFP is still located at the mitotic centrosome when Nup62-Scarlet starts to dissociate from the NPCs (−30 s, arrow). The time point of 0 s indicates the centrosome splitting which appears as 2 dots (arrowhead). Nup62 dissociation occurred approx. 33 ± 8.4 s (mean and standard deviation for n = 9) prior to centrosome duplication (see Table S3). CP75-GFP leaves the centrosome after the duplication process (40 s). Image stacks of 10 layers (z-distance of 0.5 µm) were recorded with a 10 s time lapse and only 1 focal plane is presented. Bar 5 µm.