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. 2018 Nov 19;9(12):1144. doi: 10.1038/s41419-018-1196-3

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© The Author(s) 2018

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. 2 — a Ovaries of PD11 Rps26^fl/fl/Gdf9-Cre mice and Rps26^fl/fl mice were similar in size. b, c Hematoxylin/eosin staining of the paraffin slides of ovaries showing the similar morphologies of the ovaries at PD11 from Rps26^fl/fl mice (b) and Rps26^fl/fl/Gdf9-Cre mice (c). c’ Statistical analysis of the numbers of follicles in the ovaries of Rps26^fl/fl mice (b) and Rps26^fl/fl/Gdf9-Cre mice (c). *p < 0.05, **p < 0.01 as calculated by two-tailed Student’s t-tests. d Ovaries of Rps26^fl/fl/Gdf9-Cre mice at PD14 were smaller than those of Rps26^fl/fl mice. e, f The ovarian morphology at PD14 showing that there was arrest at the pre-antral follicle stage in the ovaries of Rps26^fl/fl/Gdf9-Cre mice (f, insert) compared to Rps26^fl/fl mice (e, insert). f’ Statistical analysis of the numbers of follicles in the ovaries of Rps26^fl/fl mice (e) and Rps26^fl/fl/Gdf9-Cre mice (f). *p < 0.05, **p < 0.01 as calculated by two-tailed Student’s t-tests. g, j Ovaries of Rps26^fl/fl/Gdf9-Cre mice at PD28 (g) and PD84 (j) were smaller compared to ovaries of Rps26^fl/fl mice. h, i Ovarian morphology in Rps26^fl/fl/Gdf9-Cre mice at PD28 also showing arrest at the pre-antral follicle stage (i, insert) compared to the ovarian follicles in Rps26^fl/fl mice that had developed into pre-ovulatory and antral follicles (h, insert). i’ Statistical analysis of the numbers of follicles in the ovaries of Rps26^fl/fl mice (h) and Rps26^fl/fl/Gdf9-Cre mice (i). **p < 0.01 as calculated by two-tailed Student’s t-tests. k, l Immunohistochemistry for the oocyte marker DEAD-Box Helicase 4 (DDX4) on ovarian slides of PD84 Rps26^fl/fl/Gdf9-Cre mice (l, insert) showing no oocytes compared to those of Rps26^fl/fl mice (k, insert). l’ Statistical analysis of the numbers of follicles in the ovaries of Rps26^fl/fl mice (k) and Rps26^fl/fl/Gdf9-Cre mice (l). **p < 0.01 as calculated by two-tailed Student’s t-tests. m, n GV oocytes collected from the ovaries of Rps26^fl/fl mice (m) were larger in diameter than those from the ovaries of Rps26^fl/fl/Gdf9-Cre mice (n) at PD21–28. o, p After 12 h in vitro culture, oocyte meiosis was arrested at the GV stage in Rps26^fl/fl/Gdf9-Cre mice (p), while the oocytes of Rps26^fl/fl mice had developed into MII oocytes (o). q The quantification of the diameters of GV oocytes shown in m and n showing that oocytes collected from the ovaries of Rps26^fl/fl/Gdf9-Cre mice were significantly smaller than GV oocytes from the ovaries of Rps26^fl/fl mice. **p < 0.01 as calculated by two-tailed Student’s t-tests. r Quantification of GVBD and PBE (shown in o and p) oocytes during meiosis. The proportions of GVBD and PBE were significantly decreased in vitro in the oocytes from the ovaries of Rps26^fl/fl/Gdf9-Cre mice compared to those from Rps26^fl/fl mice. **p < 0.01 as calculated by two-tailed Student’s t-tests. s Oocyte superovulation was inhibited in Rps26^fl/fl/Gdf9-Cre mice compared to Rps26^fl/fl mice. ** p < 0.01 as calculated by two-tailed Student’s t-tests