Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Graphical description of the principle of dTAG-13 proteolysis targeting chimera-mediated endogenous protein degradation. ( B ) Graphical description of gene arrangements in REC8-FKBP12 F36V -mClover3 knockin mice. Boxes depicting genes are drawn to scale. ( C ) A representative genotyping agarose gel analysis of REC8-FKBP12 F36V -mClover3 mouse genomic DNA using oligonucleotide pairs designed to distinguish between wild-type and knockin REC8 alleles. ( D ) Representative maximum intensity projected immunofluorescence images showing that GFP antibodies specifically recognize mClover3 but not mScarlet in mClover3-NLS or mScarlet-NLS expressing prophase-arrested wild-type oocytes. ( E ) Representative maximum intensity projected immunofluorescence images of REC8 (detected with GFP antibody) and chromosomes in metaphase I-stage oocytes isolated from REC8-FKBP12 F36V -mClover3 mice. Boxes represent regions of interest magnified in lower panels.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Knock-In, Agarose Gel Electrophoresis, Immunofluorescence, Expressing, Isolation

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Representative confocal immunofluorescence microscopy images of REC8 (marked with GFP antibodies) and chromosomes (Hoechst) in metaphase I-stage oocytes of a REC8−FKBP12 F36V −mClover3 mouse. Single confocal sections spaced 1.5 µm apart are shown. b , Images from a representative high-resolution time-lapse movie of REC8−FKBP12 F36V −mClover3 (endogenous REC8) and chromosomes (SiR-5-Hoechst) showing chromosome arm-specific removal of REC8 cohesin (and its retention at centromeric regions) during anaphase of meiosis I. c , Quantification of the number of anaphase I lagging chromosomes per cell in wild-type and REC8−FKBP12 F36V −mClover3 oocytes. Data points represent individual oocytes. d , Quantification of the proportion of oocytes with zero or more anaphase I lagging chromosomes in wild-type and REC8−FKBP12 F36V −mClover3 oocytes. e , Quantification of the proportion of oocytes that completed meiosis within 16 hours of GVBD in wild-type and REC8−FKBP12 F36V −mClover3. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( c – e ). The number of analyzed oocytes is indicated in brackets and italics. GVBD, germinal vesicle breakdown; N.S., non-significant values.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Immunofluorescence, Microscopy

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Images from representative time lapse movies of chromosomes (marked with H2B-mScarlet) and meiotic spindles (marked with MAP4 microtubule-binding domain (mNeonGreen-MAP4-MTBD)) in wild-type and REC8-FKBP12 F36V -mClover3 oocytes. ( B ) Representative metaphase II chromosome spreads with centromeres labeled by ACA staining in wild-type and REC8-FKBP12 F36V -mClover3 oocytes. ( C ) The percentage of aneuploid eggs matured in vitro from wild-type or REC8-FKBP12 F36V -mClover3 oocytes. ( D ) Representative images of metaphase II-arrested eggs matured in vitro from REC8-FKBP12 F36V -mClover3 oocytes treated with DMSO or dTAG-13 for 1 h and stained with ACA and Hoechst. ( E ) Quantification of the percentage of eggs with prematurely separated sister chromatids following 1-h dTAG-13 treatment during metaphase I followed by washout and progression into meiosis II. ( F ) Representative images of embryos at various stages of preimplantation development following fertilization of DMSO- or dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. ( G ) Quantification of developmental progression from the zygote to blastocyst stage in embryos generated by fertilization of DMSO- or dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. Data are from three independent experiments. Statistical significance was assessed using Student’s t-test. The number of analyzed cells or embryos is indicated in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Binding Assay, Labeling, Staining, In Vitro, Generated

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Endogenous REC8 (labeled with GFP antibody) and metaphase I chromosomes (Hoechst) in DMSO-treated or dTAG-13-treated oocytes isolated from REC8−FKBP12 F36V −mClover3 mice. Single confocal sections spaced 1 µm apart are shown. b , Quantification of REC8 (labeled with GFP antibody) fluorescence intensity in DMSO-treated or dTAG-13-treated metaphase I oocytes isolated from REC8−FKBP12 F36V −mClover3 mice. c , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. t = 0 minutes denotes the start of live imaging experiment. d , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. e , Quantification of the proportion of cells containing single chromatids in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. f , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated or dTAG-13-treated metaphase II-arrested eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( b , d , e ) or two-way ANOVA ( f ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Labeling, Isolation, Fluorescence, In Vitro, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Western blotting analyses of REC8-FKBP12 F36V -mClover3 spermatocyte extracts treated with DMSO (control) or varying concentrations of dTAG-13. GFP antibody was used for detection of REC8 protein. Actin was used as loading control.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Western Blot, Control

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Representative confocal images of REC8 (GFP antibody) and chromosomes (Hoechst) in metaphase I-stage REC8−FKBP12 F36V −mClover3 oocytes treated with DMSO (0 minutes) or dTAG-13 for 30 minutes, 60 minutes or 90 minutes. b , Quantification of normalized REC8 fluorescence intensity in metaphase I oocytes treated with dTAG-13 for the indicated durations, fixed immediately after treatment. Fluorescence was measured on chromosomes and normalized to DMSO control. c , Representative confocal images of metaphase II chromosomes (Hoechst) and centromeres (ACA) in oocytes treated with dTAG-13 for the indicated durations during metaphase I, followed by washout and progression into meiosis II. d , Quantification of the number of prematurely separated sister chromatids per metaphase II egg after the treatment and washout scheme in c . e , Integrated analysis of the data from b and d showing the relationship between REC8 fluorescence intensity and the number of prematurely separated sister chromatids per egg. REC8 levels were measured in metaphase I oocytes immediately after treatment, and corresponding chromatid separation was quantified in metaphase II eggs after washout and meiotic progression. Chromatid separation remained minimal until REC8 levels dropped below approximately 50% of control, beyond which separation increased sharply, supporting a threshold-based model of cohesion loss. Data are from three independent experiments. Statistical comparisons were performed using Student’s t -test ( b , d ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Fluorescence, Control

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) High-resolution images of live, metaphase II-arrested wild-type mouse eggs co-expressing mClover3-MAP4-MTBD and mScarlet-tagged Gephyrin or GFP nanobodies.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Expressing

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Graphical description of nanobody-based approach for rapid degradation of endogenous REC8 in REC8−FKBP12 F36V −mClover3 oocytes. Recognition of mClover3-tagged REC8 by IgG−Fc1 fusion GFP nanobodies in TRIM21-expressing REC8−FKBP12 F36V −mClover3 eggs induces endogenous REC8 degradation via TRIM-Away and generates prematurely separated sister chromatids. b , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. t = 0 minutes denotes the start of live imaging experiment. c , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. d , Quantification of the proportion of cells containing single chromatids in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. e , Measurements of chromatid scattering volumes from time-lapse movies of Gephyrin or GFP TRIM-Away metaphase II-arrested eggs from REC8−FKBP12 F36V −mClover3 mice. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( c , d ) or two-way ANOVA ( e ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Expressing, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated, dTAG-13-treated, Cytochalasin D (CytoD)-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. t = 0 minutes denotes the start of live imaging experiment. b , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in DMSO-treated, dTAG-13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. c , Quantification of the proportion of cells containing single chromatids in DMSO-treated, dTAG13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. d , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated, dTAG-13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was assessed using Student’s t -test ( b , c ) or two-way ANOVA ( d ). The number of analyzed oocytes is provided in brackets and italics. N.S., non-significant values.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: In Vitro, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Representative confocal images of metaphase II-arrested REC8-FKBP12 F36V -mClover3 eggs treated with DMSO or dTAG-13 and immunostained with ACA (centromere marker) and HEC1 (kinetochore protein). Insets show magnified views of kinetochores. ( B ) Quantification of the proportion of oocytes exhibiting fragmented kinetochores after treatment. No significant increase in kinetochore fragmentation was observed in dTAG-13–treated REC8-FKBP12 F36V -mClover3 eggs compared to controls. ( C ) Representative images of γH2AX immunostaining in REC8-FKBP12 F36V -mClover3 metaphase II eggs treated with DMSO or dTAG-13, showing comparable levels of DNA damage marker signal. ( D ) Quantification of normalized γH2AX fluorescence intensity. No significant increase in γH2AX signal was detected in dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. ( E ) Positive control: Representative γH2AX staining of wild-type metaphase II eggs treated with etoposide, a topoisomerase II inhibitor known to induce DNA damage, confirming assay sensitivity. Data are from three independent experiments. Statistical comparisons were performed using Student’s t-test. The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Marker, Immunostaining, Fluorescence, Positive Control, Staining

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. t = 0 minutes denotes the start of live imaging experiment. b , Representative maximum intensity projected immunofluorescence images of CENP-A and chromosomes in metaphase II chromosomal spreads of DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. Boxes mark regions of interest that are 4.8× magnified. c , Quantification of CENP-A fluorescence intensities in metaphase II chromosomal spreads of DMSO-treated and IgG TRIM-Away (295 centromeres), DMSO-treated and CENP-A TRIM-Away (535 centromeres), dTAG-13-treated and IgG TRIM-Away (347 centromeres) and dTAG-13-treated and CENP-A TRIM-Away (465 centromeres) eggs from REC8−FKBP12 F36V −mClover3 mice. d , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was assessed using two-way ANOVA ( c , d ). The number of analyzed oocytes is provided in brackets and italics.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Imaging, Immunofluorescence, Fluorescence

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Quantification of the number of single chromatids per egg at 120 min of live imaging in DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8-FKBP12 F36V -mClover3 mice.

Article Snippet: The mouse strains used in this study included REC8−FKBP12 F36V −mClover3 and wild-type C57BL/6J (The Jackson Laboratory).

Techniques: Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Graphical description of the principle of dTAG-13 proteolysis targeting chimera-mediated endogenous protein degradation. ( B ) Graphical description of gene arrangements in REC8-FKBP12 F36V -mClover3 knockin mice. Boxes depicting genes are drawn to scale. ( C ) A representative genotyping agarose gel analysis of REC8-FKBP12 F36V -mClover3 mouse genomic DNA using oligonucleotide pairs designed to distinguish between wild-type and knockin REC8 alleles. ( D ) Representative maximum intensity projected immunofluorescence images showing that GFP antibodies specifically recognize mClover3 but not mScarlet in mClover3-NLS or mScarlet-NLS expressing prophase-arrested wild-type oocytes. ( E ) Representative maximum intensity projected immunofluorescence images of REC8 (detected with GFP antibody) and chromosomes in metaphase I-stage oocytes isolated from REC8-FKBP12 F36V -mClover3 mice. Boxes represent regions of interest magnified in lower panels.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Knock-In, Agarose Gel Electrophoresis, Immunofluorescence, Expressing, Isolation

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Representative confocal immunofluorescence microscopy images of REC8 (marked with GFP antibodies) and chromosomes (Hoechst) in metaphase I-stage oocytes of a REC8−FKBP12 F36V −mClover3 mouse. Single confocal sections spaced 1.5 µm apart are shown. b , Images from a representative high-resolution time-lapse movie of REC8−FKBP12 F36V −mClover3 (endogenous REC8) and chromosomes (SiR-5-Hoechst) showing chromosome arm-specific removal of REC8 cohesin (and its retention at centromeric regions) during anaphase of meiosis I. c , Quantification of the number of anaphase I lagging chromosomes per cell in wild-type and REC8−FKBP12 F36V −mClover3 oocytes. Data points represent individual oocytes. d , Quantification of the proportion of oocytes with zero or more anaphase I lagging chromosomes in wild-type and REC8−FKBP12 F36V −mClover3 oocytes. e , Quantification of the proportion of oocytes that completed meiosis within 16 hours of GVBD in wild-type and REC8−FKBP12 F36V −mClover3. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( c – e ). The number of analyzed oocytes is indicated in brackets and italics. GVBD, germinal vesicle breakdown; N.S., non-significant values.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Immunofluorescence, Microscopy

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Images from representative time lapse movies of chromosomes (marked with H2B-mScarlet) and meiotic spindles (marked with MAP4 microtubule-binding domain (mNeonGreen-MAP4-MTBD)) in wild-type and REC8-FKBP12 F36V -mClover3 oocytes. ( B ) Representative metaphase II chromosome spreads with centromeres labeled by ACA staining in wild-type and REC8-FKBP12 F36V -mClover3 oocytes. ( C ) The percentage of aneuploid eggs matured in vitro from wild-type or REC8-FKBP12 F36V -mClover3 oocytes. ( D ) Representative images of metaphase II-arrested eggs matured in vitro from REC8-FKBP12 F36V -mClover3 oocytes treated with DMSO or dTAG-13 for 1 h and stained with ACA and Hoechst. ( E ) Quantification of the percentage of eggs with prematurely separated sister chromatids following 1-h dTAG-13 treatment during metaphase I followed by washout and progression into meiosis II. ( F ) Representative images of embryos at various stages of preimplantation development following fertilization of DMSO- or dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. ( G ) Quantification of developmental progression from the zygote to blastocyst stage in embryos generated by fertilization of DMSO- or dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. Data are from three independent experiments. Statistical significance was assessed using Student’s t-test. The number of analyzed cells or embryos is indicated in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Binding Assay, Labeling, Staining, In Vitro, Generated

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Endogenous REC8 (labeled with GFP antibody) and metaphase I chromosomes (Hoechst) in DMSO-treated or dTAG-13-treated oocytes isolated from REC8−FKBP12 F36V −mClover3 mice. Single confocal sections spaced 1 µm apart are shown. b , Quantification of REC8 (labeled with GFP antibody) fluorescence intensity in DMSO-treated or dTAG-13-treated metaphase I oocytes isolated from REC8−FKBP12 F36V −mClover3 mice. c , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. t = 0 minutes denotes the start of live imaging experiment. d , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. e , Quantification of the proportion of cells containing single chromatids in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. f , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated or dTAG-13-treated metaphase II-arrested eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( b , d , e ) or two-way ANOVA ( f ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Labeling, Isolation, Fluorescence, In Vitro, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Western blotting analyses of REC8-FKBP12 F36V -mClover3 spermatocyte extracts treated with DMSO (control) or varying concentrations of dTAG-13. GFP antibody was used for detection of REC8 protein. Actin was used as loading control.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Western Blot, Control

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Representative confocal images of REC8 (GFP antibody) and chromosomes (Hoechst) in metaphase I-stage REC8−FKBP12 F36V −mClover3 oocytes treated with DMSO (0 minutes) or dTAG-13 for 30 minutes, 60 minutes or 90 minutes. b , Quantification of normalized REC8 fluorescence intensity in metaphase I oocytes treated with dTAG-13 for the indicated durations, fixed immediately after treatment. Fluorescence was measured on chromosomes and normalized to DMSO control. c , Representative confocal images of metaphase II chromosomes (Hoechst) and centromeres (ACA) in oocytes treated with dTAG-13 for the indicated durations during metaphase I, followed by washout and progression into meiosis II. d , Quantification of the number of prematurely separated sister chromatids per metaphase II egg after the treatment and washout scheme in c . e , Integrated analysis of the data from b and d showing the relationship between REC8 fluorescence intensity and the number of prematurely separated sister chromatids per egg. REC8 levels were measured in metaphase I oocytes immediately after treatment, and corresponding chromatid separation was quantified in metaphase II eggs after washout and meiotic progression. Chromatid separation remained minimal until REC8 levels dropped below approximately 50% of control, beyond which separation increased sharply, supporting a threshold-based model of cohesion loss. Data are from three independent experiments. Statistical comparisons were performed using Student’s t -test ( b , d ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Fluorescence, Control

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) High-resolution images of live, metaphase II-arrested wild-type mouse eggs co-expressing mClover3-MAP4-MTBD and mScarlet-tagged Gephyrin or GFP nanobodies.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Expressing

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Graphical description of nanobody-based approach for rapid degradation of endogenous REC8 in REC8−FKBP12 F36V −mClover3 oocytes. Recognition of mClover3-tagged REC8 by IgG−Fc1 fusion GFP nanobodies in TRIM21-expressing REC8−FKBP12 F36V −mClover3 eggs induces endogenous REC8 degradation via TRIM-Away and generates prematurely separated sister chromatids. b , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. t = 0 minutes denotes the start of live imaging experiment. c , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. d , Quantification of the proportion of cells containing single chromatids in Gephyrin or GFP TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. e , Measurements of chromatid scattering volumes from time-lapse movies of Gephyrin or GFP TRIM-Away metaphase II-arrested eggs from REC8−FKBP12 F36V −mClover3 mice. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was evaluated using Student’s t -test ( c , d ) or two-way ANOVA ( e ). The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Expressing, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated, dTAG-13-treated, Cytochalasin D (CytoD)-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. t = 0 minutes denotes the start of live imaging experiment. b , Quantification of the number of single chromatids per egg at 90 minutes of live imaging in DMSO-treated, dTAG-13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. c , Quantification of the proportion of cells containing single chromatids in DMSO-treated, dTAG13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. d , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated, dTAG-13-treated, CytoD-treated or dTAG-13-treated and CytoD-treated eggs matured in vitro from REC8−FKBP12 F36V −mClover3 oocytes. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was assessed using Student’s t -test ( b , c ) or two-way ANOVA ( d ). The number of analyzed oocytes is provided in brackets and italics. N.S., non-significant values.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: In Vitro, Imaging

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Representative confocal images of metaphase II-arrested REC8-FKBP12 F36V -mClover3 eggs treated with DMSO or dTAG-13 and immunostained with ACA (centromere marker) and HEC1 (kinetochore protein). Insets show magnified views of kinetochores. ( B ) Quantification of the proportion of oocytes exhibiting fragmented kinetochores after treatment. No significant increase in kinetochore fragmentation was observed in dTAG-13–treated REC8-FKBP12 F36V -mClover3 eggs compared to controls. ( C ) Representative images of γH2AX immunostaining in REC8-FKBP12 F36V -mClover3 metaphase II eggs treated with DMSO or dTAG-13, showing comparable levels of DNA damage marker signal. ( D ) Quantification of normalized γH2AX fluorescence intensity. No significant increase in γH2AX signal was detected in dTAG-13-treated REC8-FKBP12 F36V -mClover3 eggs. ( E ) Positive control: Representative γH2AX staining of wild-type metaphase II eggs treated with etoposide, a topoisomerase II inhibitor known to induce DNA damage, confirming assay sensitivity. Data are from three independent experiments. Statistical comparisons were performed using Student’s t-test. The number of analyzed oocytes is indicated in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Marker, Immunostaining, Fluorescence, Positive Control, Staining

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: a , Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. t = 0 minutes denotes the start of live imaging experiment. b , Representative maximum intensity projected immunofluorescence images of CENP-A and chromosomes in metaphase II chromosomal spreads of DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. Boxes mark regions of interest that are 4.8× magnified. c , Quantification of CENP-A fluorescence intensities in metaphase II chromosomal spreads of DMSO-treated and IgG TRIM-Away (295 centromeres), DMSO-treated and CENP-A TRIM-Away (535 centromeres), dTAG-13-treated and IgG TRIM-Away (347 centromeres) and dTAG-13-treated and CENP-A TRIM-Away (465 centromeres) eggs from REC8−FKBP12 F36V −mClover3 mice. d , Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8−FKBP12 F36V −mClover3 mice. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was assessed using two-way ANOVA ( c , d ). The number of analyzed oocytes is provided in brackets and italics.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Imaging, Immunofluorescence, Fluorescence

Journal: Nature Aging

Article Title: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy

doi: 10.1038/s43587-025-00997-w

Figure Lengend Snippet: ( A ) Quantification of the number of single chromatids per egg at 120 min of live imaging in DMSO-treated and IgG TRIM-Away, DMSO-treated and CENP-A TRIM-Away, dTAG-13-treated and IgG TRIM-Away or dTAG-13-treated and CENP-A TRIM-Away eggs from REC8-FKBP12 F36V -mClover3 mice.

Article Snippet: REC8−FKBP12 F36V −mClover3 mice are in the process of being deposited to The Jackson Laboratory for sharing with the wider research community (expected availability in 2026).

Techniques: Imaging