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. 2025 Dec 12;17:720. doi: 10.1038/s41467-025-67476-z

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© The Author(s) 2025

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 4 — a Immunofluorescence images oocytes from follicles at different growth stages on mouse ovary sections. b, c Automated quantification of total intensity of pS2 and RPB1 in oocytes from follicles at different growth stages on mouse ovary sections. Compared to primordial follicle, P_adjusted(primary follicle-pS2)=0.60178; P_adjusted(primary follicle-RPB1) = 0.97547; P_adjusted(secondary follicle-pS2)=0.99954; P_adjusted(secondary follicle-RPB1) = 0.00911; P_adjusted(antral follicle-pS2)=0; P_adjusted(antral follicle-RPB1) = 0. d Immunofluorescence images of oocytes harvested from mouse secondary follicles cultured for different days. e–h Automated quantification of total intensity of pS2, RPB1, chromatin-bound RPB1 and chromatin volume in growing oocytes harvested from mouse secondary follicles cultured for different days. Compared to D14 + 3, P_adjusted(D14 + 5-pS2)=1.55805E-4; P_adjusted(D14 + 5-RPB1) = 1.21251E-6; P_adjusted(D14 + 5-chromatin-bound RPB1) = 9.15881E-8; P_adjusted(D14 + 5-chromatin volume)=0.14783; P_adjusted(D14 + 7-pS2)=0.86434; P_adjusted(D14 + 7-RPB1) = 0.16675; P_adjusted(D14 + 7-chromatin-bound RPB1) = 1.39079E-5; P_adjusted(D14 + 7-chromatin volume)=0.20583; P_adjusted(D14 + 9-pS2)=5.37076E-4; P_adjusted(D14 + 9-RPB1) = 0.06188; P_adjusted(D14 + 9-chromatin-bound RPB1) = 0.9796; P_adjusted(D14 + 9-chromatin volume)=0. i Immunofluorescence images of oocytes harvested from mouse secondary follicles treated with CB-5083, MG132 and flavopiridol on D14 + 7 for 2 days. Yellow arrowheads highlight the nucleolus. j Automated quantification of total intensity of RPB1 in oocytes harvested from mouse secondary follicles treated with CB-5083, MG132 or flavopiridol(Flavo.) on D14 + 7 for 2 days. Compared to DMSO, P_adjusted(CB-5083) = 0; P_adjusted(MG132) = 1.07948E-6; P_adjusted(flavopiridol)=0.77002. k Manual scoring of chromatin configurations in oocytes harvested from mouse secondary follicles treated with CB-5083 (two-tailed Fisher’s exact test, p = 0.0169), MG132 (two-tailed Fisher’s exact test, p = 0.0569) or flavopiridol. l Immunofluorescence images of NSN and SN oocytes from wildtype and Ubb cKO mice. m Manual scoring of chromatin configurations in oocytes from wildtype and Ubb cKO mice (two-tailed Fisher’s exact test, p = 0.1296). *p < 0.05. ****p < 0.0001. N.S., not significant. Scatter intervals are presented as mean values +/- S.D. Statistical significance is based on one-way ANOVA followed by post-hoc Tukey’s multiple comparison test (b, c, e–h, j). The number of oocytes analyzed is specified in italics. Scale bars are 5 µm. Source data are provided as a Source Data file.