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. 2019 Mar 26;9:5167. doi: 10.1038/s41598-019-41610-6

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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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Meiotic maturation following forskolin (FSK) and 3-isobutyl-1-methylxanthine (IBMX) treatment. Change in nuclear status over time was assessed following FSK and IBMX treatment. During in vitro maturation, 20 oocytes per treatment from 3 independent experiments were fixed at several time points and stained to assess the time-dependent change of nuclear maturation. A total of 1080 oocytes were examined. Time 0 represents the start of maturation culture. Open circle: germinal vesicle breakdown (GVBD) in control, closed circle: GVBD following FSK and IBMX treatment, open square: metaphase II oocytes in control, closed square: metaphase II oocytes following FSK and IBMX treatment.