Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a Relative mRNA levels of Kitl in cultured PD10 ovarian cells after securinine treatment (≥ 5 independent experiments) determined by RT-qPCR. Ovarian cells isolated from PD10 mouse ovaries were cultured overnight in DMEM/F-12 medium supplemented with 10% fetal bovine serum (FBS), followed by serum starvation in FBS-free medium for 18 h. The starved cells were then treated with various concentrations of securinine for 24 h. Data are shown as the mean ± SEM. b Morphology of PD7 ovaries after 6 days of culture with or without 20 μM securinine. Black arrows indicate primordial follicles, black arrowheads indicate primary follicles, and red arrowhead indicate secondary follicles. Hematoxylin staining indicates the nuclei. Scale bars: 50 μm. c Ovarian follicles at different developmental stages were counted across all serial sections of whole ovaries ( n = 6 ovaries for the control group; n = 7 ovaries for the securinine-treated group). Data are shown as the mean ± SEM. d Comparison of FOXO3 (green) subcellular localization in oocytes. Securinine promoted the nuclear export of FOXO3 into the cytoplasm of activated follicle oocytes (red arrowheads), which was in contrast to the localization of FOXO3 in the nuclei of primordial follicle oocytes (white arrows). Scale bars: 50 µm. The experiment was repeated at least three times, and representative images are shown. Data were analyzed by a two-tailed unpaired Student’s t test without adjustments. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ns, not significant. Source data are provided as a Source Data file, including exact P- values.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: Cell Culture, Quantitative RT-PCR, Isolation, Staining, Control, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a I: An illustration of the intraovarian injection of securinine in aged ovaries of 10–12-month-old mice. For each mouse, one lateral ovary was injected with 20 µL of securinine (1 mM), while the contralateral ovary was injected with 20 µL hyaluronic acid alone (carrier) and served as the control. The injected ovaries were collected 10 days after the intraovarian injection. II : An illustration of a fertility test in 10-month-old C57BL/6J female mice subjected to intraovarian injection of securinine. b The histological morphology of control and securinine-treated ovaries at 10 days after injection. Red arrowheads indicate secondary follicles, and red arrows indicate antral follicles. Hematoxylin staining indicates nuclei. Scale bars: 100 μm. c The distribution of follicles in ovaries treated with or without securinine at 10 days after injection. Ovarian follicles at various developmental stages, including primordial, primary, secondary, and antral follicles and corpora lutea, were counted in all sections of the ovary, and a total of 22 ovaries were analyzed per group. Data are shown as the mean ± SEM. d Cumulative numbers of pups per group during the observation period. The cumulative numbers of pups observed in the securinine-treated group (red line) were greater compared to the control group (black line), n = 12 female parental mice per group. e The weights of the pups at 8 weeks of age, n = 18 pups per group. Data are shown as the mean ± SEM. Data were analyzed by a two-tailed unpaired Student’s t test without adjustments. ∗ P < 0.05; ∗∗ P < 0.01; ns, not significant. Source data are provided as a Source Data file, including exact P- values.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: Injection, Control, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a Schematic diagram of the scRNA-seq procedure. Whole ovaries were collected from 10–12-month-old C57BL/6J mice 2 days after intraovarian injection and were processed for 10 × Genomics 3′ scRNA-seq, with 4 ovaries (from 4 mice) pooled per treatment group to generate each sequencing library. b UMAP plot of ovarian cells. Clustering analysis revealed 15 distinct ovarian cell populations. c The percentages of cells in broad cell type categories. d Network diagrams illustrating the changes in the number of ligand–receptor interaction events across different cell types in the ovaries after intraovarian injection of securinine. e Network diagrams illustrating the changes in the interaction strength of ligand–receptor interaction events across different cell types in the ovaries after intraovarian injection of securinine. f GO enrichment of changed interaction pairs in stromal cells and granulosa cells. g GO enrichment of differentially expressed ECM-related genes in stromal cells and granulosa cells.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: Injection, Sequencing

Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a Immunofluorescence staining of COL1A1 in ovarian tissues from 10-month-old mice. Representative images show the spatial distribution of stromal cells in relation to secondary follicles (white arrowheads). GC indicates granulosa cells. Scale bars: 50 µm. The experiment was repeated at least three times, and representative images are shown. b Immunofluorescence staining of DCN in ovarian tissues from 10-month-old mice. Representative images show the spatial distribution of stromal cells adjacent to both secondary follicles (white arrowheads) and antral follicles (white arrows). Scale bars: 50 µm. The experiment was repeated at least three times, and representative images are shown. c Reduced interaction of ligand–receptor pairs from stromal cells to granulosa cells after securinine injection. d Relative mRNA levels of Col1a2 , Col1a1 , and Col6a3 genes in cultured PD10 ovarian cells after securinine treatment (4 independent experiments) determined by RT-qPCR. Securinine treatment significantly reduced the expression of these ECM-related genes. Data are shown as the mean ± SEM. e Comparison of the morphology of control and doxycycline-treated ovaries. Black arrows indicate primordial follicles; black arrowheads indicate primary follicles, and red arrowhead indicates a secondary follicle. Hematoxylin staining indicates nuclei. Scale bars: 50 μm. f Ovarian follicles at different developmental stages were counted across all serial sections of whole ovaries ( n = 6 ovaries for the control group; n = 7 ovaries for the doxycycline-treated group). Data are shown as the mean ± SEM. Data were analyzed by a two-tailed unpaired Student’s t test without adjustments. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ns, not significant. Source data are provided as a Source Data file, including exact P- values.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: Immunofluorescence, Staining, Injection, Cell Culture, Quantitative RT-PCR, Expressing, Comparison, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a An illustration of the in vitro treatment of cynomolgus macaque ovarian cortical tissues with securinine followed by transplantation under the kidney capsules of nude mice. Ovarian fragments were carefully cut into small cubes, treated with or without securinine (20 μM) for 24 h, and transplanted under the kidney capsules of BALB/c nude mice. Created in BioRender. Liu, D. (2026) https://BioRender.com/b1odufa . b The histological morphology of fresh ovarian tissues from cynomolgus macaques. Black arrows indicate primordial follicles. Hematoxylin staining indicates the nuclei. Scale bars: 100 μm. c The histological morphology of control and securinine-treated ovarian tissues at 23 days after engraftment. Ovarian tissues pretreated with securinine showed more antral follicles (red arrows) and corpora lutea (CL, yellow arrows) as compared to the control tissues, which predominantly contained primordial follicles (black arrows). Hematoxylin staining indicates the nuclei. Scale bars: 100 μm. d The distribution of follicles in ovarian tissues treated with or without securinine at 23 days after transplantation ( n = 10 pieces of ovarian tissues from 10 cynomolgus macaques). Data are shown as the mean ± SEM. Data were analyzed by a two-tailed unpaired Student’s t test without adjustments. ∗∗ P < 0.01; ns, not significant. Source data are provided as a Source Data file, including exact P- values.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: In Vitro, Transplantation Assay, Capsules, Staining, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Intraovarian injection of securinine stimulates the growth of small ovarian follicles by reducing stromal suppression

doi: 10.1038/s41467-026-71691-7

Figure Lengend Snippet: a Schematic illustration of the intraovarian injection of securinine into cynomolgus macaques. Each animal received a unilateral injection into one ovary with 300 μL of a 6 mM securinine solution suspended in 10 mg/mL hyaluronic acid carrier. The contralateral ovary was injected with hyaluronic acid alone as an internal control. Created in BioRender. Liu, D. (2026) https://BioRender.com/awud1u2 . b B-mode ultrasonography showing ovarian position (red ovals) and follicular development just prior to intraovarian injection and at 7 and 14 days following securinine injection. Red arrows indicate possible antral follicles, while white arrows indicate a corpus luteum (CL). Scale bars: 5 mm. c Images showing the morphology of securinine-injected and control ovaries at 14 days after injection. Red arrows indicate possible antral follicles observed in the securinine-injected ovary. Scale bars: 1 cm. d Histological analysis showed that at 14 days after injection the control ovary mainly contained primordial (black arrows) and secondary follicle (red arrowhead), whereas the securinine-injected ovary contained antral follicles (red arrows). Scale bars: 100 µm. e Immunofluorescence staining of DDX4 (red) showed antral follicles (red arrow) in securinine-injected ovaries compared to the control ovary. Scale bars: 100 µm. The experiment was repeated at least three times, and representative images are shown. f Schematic diagram illustrating how securinine promotes follicular development in aged ovaries.

Article Snippet: The concentration of securinine (#HY-N2079, MedChemExpress, Monmouth Junction, NJ, USA) was 20 μM for in vitro culture.

Techniques: Injection, Control, Immunofluorescence, Staining