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. 2022 Feb 17;13:914. doi: 10.1038/s41467-022-28611-2

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© The Author(s) 2022

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. 3 — a Experimental design of Ab4B19 treatment and analysis in NA-POF model. b BDNF concentration in ovaries at different ages, from 2 months to 14 months. (N = 6 mice per group). c Pharmacodynamic analysis showing the number of ovulated oocytes per mouse treated with increasing doses of Ab4B19 from 0.1 mg/kg to 1 mg/kg. (N = 8 mice per group). d–j Aged mice of 12–13 months old were treated with Ab4b19 or IgG (1 mg/kg) every 4 days through tail vein injection. After 16-day (four doses) treatment, ovaries and serum were collected for analysis. d Representative estrous cycles of NA-POF mice treated with Ab4B19 (left) or normal IgG (right). P proestrus stage, E estrus stage, M metestrus stage, D diestrus stage. e Effect of Ab4B19 on time spent in various estrous stages. Stages of the estrous cycle were determined by microscopic analysis of the predominant cell types in the vaginal smears. (N = 8 mice per group). f Representative images of H&E staining of ovaries from NA-POF mice treated with Ab4B19 or vehicle. Red arrow: atretic ovarian follicle; Black arrow: antral ovarian follicle; Asterisk: corpora lutea. Scale bars, 200 μm. g, h Quantitative analysis of the primordial follicles, pre-antral follicles, antral follicles, and corpora lutea, as well as the atretic follicles. (N = 6 ovaries in vehicle group and 8 ovaries in Ab4B19 group). The serum levels of estrogen (i) and FSH (j) were measured by ELISA (N = 7 mice in vehicle group and 8 mice in Ab4B19 group). k Representative Western blot and quantitative analyses of FSHR and AMH expression. β-actin was used as a loading control. N = 4 mice in each condition, with the same samples repeated at least twice (n = 8). All experiments were repeated at least three times. The data were shown as the mean ± SEM with individual values. Statistical analyses were carried out using one-way ANOVA followed by the Dunnett’s multiple comparisons test for (b, c), or two-tailed student’s t-test (e, g–k), respectively (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns not significant). Source data are provided as a Source Data file.