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. 2021 Jul 14;12:4322. doi: 10.1038/s41467-021-24554-2

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© The Author(s) 2021

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. 1 — a Graph showing the mean destruction profiles of VFP-tagged securin FL (magenta trace, n = 25) and cyclin B1 FL (blue trace, n = 62) alongside separase activity as determined by a separase activity biosensor (mCherry/eGFP ratio, n = 21) in MI mouse oocytes relative to PB1 extrusion. Error bars ± SEM. Representative confocal images show a maturing oocyte expressing Map4-GFP (microtubules in green) and incubated with SiR-DNA (DNA in red) at times relative to the x axis of the graph. Scale bar = 10 µm. b Schematic diagram of an H2B-mCherry-Scc1-eGFP separase activity biosensor. c Representative time-lapse images of an oocyte expressing the separase activity biosensor in Fig. 1b, imaged every 10 minutes at the times indicated relative to PB1 extrusion. eGFP (green), mCherry (red) and merged fluorescence + bright-field (BF) images are shown. Scale bar = 10 µm. eGFP and mCherry images correspond to the areas marked by the white dotted lines in the merge + BF images. Images displayed in a and c are representative of between 10 and 20 oocytes from experiments repeated at least three times.