MATR3 is essential for oocyte growth and maturation quality through a dual molecular mechanism
- State Key Laboratory of Animal Biotech Breeding, College of Biological Sciences, China Agricultural University, China
- Department of Physiology and Pathophysiology, National Key Discipline of Cell Biology, School of Basic Medicine, Fourth Military Medical University, China
- College of Bioscience and Resources Environment, Beijing University of Agriculture, China
- Department of Gynecology, General Hospital of Ningxia Medical University, China
- Reproductive Medicine Center, General Hospital of Ningxia Medical University, China
- Institute of Medical Sciences, General Hospital of Ningxia Medical University, China
- Transgenic Animal Center, National Institute of Biological Sciences, China
Figures
Figure 1 with 3 supplements
MATR3 in growing oocyte regulates female reproduction in mice.
(A) Immunofluorescence showing MATR3 levels in mouse oocytes. (B) Western blot results showing MATR3 expression in mouse oocytes. Total proteins from 200 oocytes were loaded in each lane. GAPDH served as a loading control. (C) Relative expression level of MATR3 to GAPDH. (D) Immunofluorescence showing MATR3 levels in human oocytes. n=11. (E) Representative image of follicles and oocytes from negative control (NC) and si-Matr3. (F) Immunohistochemistry results showing MATR3 expression in oocytes. (G) Cumulative number of pups from Ctrl (n=6) and cKO (n=3) female. (H) Percentage of antral follicles in E. n≥33 per group. (I) MII ratio of oocytes collected from E. n≥33 per group. Scale bars: 20 μm in (A), (B), and (C), 40 μm in (E) and (F). Data are represented as mean ± SD. ***p<0.001, **p<0.01, *p<0.05, n.s., not significant.
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Figure 1—source data 1
PDF file containing original western blots for Figure 1B, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-data1-v1.zip
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Figure 1—source data 2
Original files for western blot analysis displayed in Figure 1B.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-data2-v1.zip
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Figure 1—source data 3
Source data for Figure 1C, G, H and I.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-data3-v1.xlsx
Figure 1—figure supplement 1
The expression pattern of MATR3 in ovaries of mice and porcine.
(A) Immunofluorescence results showing MATR3 location in mouse ovaries. Mouse ovaries were stained for MATR3 (green) at postnatal day 23 (PD23). The nuclei were dyed with a Hoechst counterstain (blue). Scale bar: 25 μm. (B) Immunohistochemistry results showing MATR3 location in porcine ovaries. MATR3 was mainly localized to the nuclei of oocytes in either mouse or porcine. Scale bar: 50 μm. (C) Western blotting was used to detect the changes of MATR3 in GCs of porcine during the growth of follicles under physiological conditions. The results showed that MATR3 was highly expressed in the GCs of small secondary follicles, n=3. (D) Western blotting was used to detect the changes of MATR3 in GCs of porcine with the duration of LH. The results showed that MATR3 increased with the duration of LH, n=3.
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Figure 1—figure supplement 1—source data 1
PDF file containing original western blots for Figure 1—figure supplement 1C, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp1-data1-v1.zip
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Figure 1—figure supplement 1—source data 2
Original files for western blot analysis displayed in Figure 1—figure supplement 1C.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp1-data2-v1.zip
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Figure 1—figure supplement 1—source data 3
PDF file containing original western blots for Figure 1—figure supplement 1D, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp1-data3-v1.zip
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Figure 1—figure supplement 1—source data 4
Original files for western blot analysis displayed in Figure 1—figure supplement 1D.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp1-data4-v1.zip
Figure 1—figure supplement 2
Construction of in vitro knockdown model, knockdown efficiency, and phenotypes.
(A) Using the method of in vitro addition of follicle-stimulating hormone (FSH), secondary follicles with a diameter of 150–200 μm were continuously cultured. The follicles could develop to the pre-ovulatory follicle stage, and oocytes with the ability to resume meiosis could be ovulated 16 hr after the addition of LH. Scale bar: 100 μm. (B) After the 3T3 cell line was transfected with negative control (NC) and Matr3 siRNA, respectively, for 48 hr, western blot was used to detect the protein level of MATR3 in the cells, with n=3. β-Actin was used as an internal reference to normalize the protein loading level. (C) The statistical results of the gray scale scanning values of the MATR3 protein bands in (B). The statistical results are expressed as mean ± SD. (D) The knockdown efficiency was detected by immunofluorescence technique. Green fluorescence: MATR3; blue fluorescence: Hoechst (nucleus). Scale bar: 25 μm. (E) Isolate the somatic cells and oocytes of the follicles that have been continuously cultured for 5 days. Detect the changes in the levels of proteins related to the proliferation and differentiation of granulosa cells by western blotting. n=3.
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Figure 1—figure supplement 2—source data 1
PDF file containing original western blots for Figure 1—figure supplement 2B, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp2-data1-v1.zip
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Figure 1—figure supplement 2—source data 2
Original files for western blot analysis displayed in Figure 1—figure supplement 2B.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp2-data2-v1.zip
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Figure 1—figure supplement 2—source data 3
PDF file containing original western blots for Figure 1—figure supplement 2E, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp2-data3-v1.zip
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Figure 1—figure supplement 2—source data 4
Original files for western blot analysis displayed in Figure 1—figure supplement 2E.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp2-data4-v1.zip
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Figure 1—figure supplement 2—source data 5
Source data for Figure 1—figure supplement 2C.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp2-data5-v1.xlsx
Figure 1—figure supplement 3
Strategies for constructing Matr3 oocyte-specific knockout mice and detection of related phenotypes.
(A) Using PCR to identify the genotypes of mice, with DL2000 as the DNA marker. After the insertion of LoxP, the size of the band is 231 bp, the size of the wild-type (WT) band is 192 bp, and the size of the Cre is 500 bp. (B) Construction strategy for Matr3 knockout model in oocyte via Gdf9-Cre. ATG: transcription start site; red arrow: LoxP insertion site; numbers 1–15: exon regions 1–15 of Matr3. (C) Immunofluorescence results showing MATR3 location in oocytes from Ctrl and cKO mouse (postnatal day 14 [PD14]). Oocytes were stained for MATR3 (green). The nuclei were dyed with a Hoechst counterstain (blue). Scale bar: 50 μm.
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Figure 1—figure supplement 3—source data 1
PDF file containing original blots for Figure 1—figure supplement 3A, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp3-data1-v1.zip
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Figure 1—figure supplement 3—source data 2
Original files for blot analysis displayed in Figure 1—figure supplement 3A.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig1-figsupp3-data2-v1.zip
Figure 2 with 2 supplements
MATR3 was required to support oocyte growth.
(A) Hematoxylin staining of postnatal day 23 (PD23) ovaries after gonadotropins injection. (B) Number of follicles in PD23 ovaries after gonadotropins injection. n=3. (C) Morphology and number of oocytes collected from oviducts of PD23 mice after superovulation. n=3. (D) Morphology of oocytes derived from PD23 mice primed with pregnant mare serum gonadotropin (PMSG) for 48 hr. n=3. (E, F) Diameter of oocytes derived from PD23 mice primed with PMSG for 48 hr. n=40. (G) Non-surrounded nucleolus (NSN) and surrounded nucleolus (SN) ratio of oocytes derived from PD23 mice primed with PMSG for 48 hr. n=3. (H) MII ratio of oocytes derived from PD23 mice primed with PMSG for 48 hr. n(Ctrl)≥38, n(cKO)≥91. (I) Hematoxylin staining of PD7, PD14, PD23, PD35 ovaries. n=3. (J) Number of total follicles in PD7, PD14, PD23, PD35 ovaries. n=3. (K) Cell proliferation indicated by Ki67-positive GCs in mice ovaries. (L) Statistics of antral follicle (AF)/secondary follicle (SF) in PD23 and PD35 ovaries. n=3. (M) Number of SFs in PD7, PD14, PD23, PD35 ovaries. n=3. Scale bars: 40 μm in (C) and (E), 120 μm in (A), (I), and (K). Data are represented as mean ± SD. ***p<0.001, **p<0.01, *p<0.05, n.s., not significant.
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Figure 2—source data 1
Source data for Figure 2B, D, F–H, J, L and M.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Detection of Matr3 oocyte-specific knockout mice phenotypes.
(A) Morphology of oocytes derived from postnatal day 14 (PD14) mice. Scale bar: 50 μm. (B) Diameter of oocytes derived from PD14 mice. n=50. (C) Number of follicles in PD7 ovaries. PrF: primordial follicle; GF: growing follicle; TF: total follicle. (D) Quantification of the ratio of Ki-67-positive granulosa cells within individual follicles. n=20. Data are represented as mean ± SD. ***p<0.001, n.s., not significant.
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Figure 2—figure supplement 1—source data 1
Source data for Figure 2—figure supplement 1C–E.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig2-figsupp1-data1-v1.xlsx
Figure 2—figure supplement 2
The Matr3 floxed allele induces a mild hypomorphic effect.
(A) Relative mRNA expression levels of Matr3 in mice of different genotypes. n=3. (B) Representative images of oocytes derived from postnatal day 23 (PD23) mice primed with pregnant mare serum gonadotropin (PMSG) for 48 hr. Scale bar: 50 μm. (C) MII ratio of oocytes derived from PD23 mice primed with PMSG for 48 hr. n (wild-type)>50, n (Matr3flox/flox)>50. Data are represented as mean ± SD. *p<0.05, **p<0.01.
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Figure 2—figure supplement 2—source data 1
Source data for Figure 2—figure supplement 2A, C.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig2-figsupp2-data1-v1.xlsx
Figure 3 with 3 supplements
Matr3 knockdown results in the reduction of transcriptional activity.
(A) Volcano plot shows differentially expressed genes (DEGs) (upregulated, red; downregulated, blue) in si-Matr3 oocytes compared to the negative control (NC). (B) Key gene ontology (GO) enrichment of all DEGs in GOs. (C) Top 10 terms of downregulated DEGs in GOs. (D) 5-Ethynyl uridine (EU) staining (green) in GOs collected from NC and si-Matr3. n ≥20. (E) Quantification of the mean fluorescence intensity of EU in oocytes. (F) RT-qPCR results showing Tbpl2 mRNA level in GOs. (G) Western blotting results showing RNAPII protein level in GOs. (H) H3K9me1 (red), H3K9me3 (red), H3K27me3 (red), H3K4me3 (green), staining in GOs collected from NC and si-Matr3. n ≥20. (I) H3K9me2 staining (red) in GOs collected from NC and si-Matr3. n ≥37. (J) Quantification of the mean fluorescence intensity of H3K9me2 in oocytes. Scale bar: 20 μm. Data are represented as mean ± SD. ***p<0.001, **p<0.01.
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Figure 3—source data 1
PDF file containing original western blots for Figure 3G, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig3-data1-v1.zip
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Figure 3—source data 2
Original files for western blot analysis displayed in Figure 3G.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig3-data2-v1.zip
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Figure 3—source data 3
Source data for Figure 3B, C, E–G and J.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig3-data3-v1.xlsx
Figure 3—figure supplement 1
The spatiotemporal-specific localization of MATR3 is adapted to the transcriptional activity of oocytes.
Immunofluorescence results showing the relationship between the localization of MATR3 in oocytes at different growth states and the transcriptional activity of oocytes. Red: MATR3; green: 5-ethynyl uridine (EU) (newly synthesized mRNA); blue: Hoechst (cell nucleus). Scale bar: 25 μm.
Figure 3—figure supplement 2
Parallelism analysis of single-cell RNA-sequencing (scRNA-seq) samples.
(A) PCA shows that the gene ontology (GO) in the negative control (NC) group and the si-Matr3 group form two subgroups. (B) Clustering heatmap analysis demonstrates the global changes in the expression levels of differentially expressed genes in the GOs of the NC group and the si-Matr3 group based on the results of scRNA-seq data. The horizontal axis represents the types of sample replicates, and the vertical axis represents different genes.
Figure 3—figure supplement 3
The histone methylation level of oocytes after Matr3 knockdown in vitro.
The statistical results of the fluorescence intensity of histone methylation, with n (H3K9me1)=20, n (H3K9me3)=40, n (H3K27me3)=18, and n (H3K4me3)=20. The statistical results are presented as mean ± SD.
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Figure 3—figure supplement 3—source data 1
Source data for Figure 3—figure supplement 3.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig3-figsupp3-data1-v1.xlsx
Figure 4
MATR3 controls oocyte growth by regulating GDF9 level.
(A) RT-qPCR results showing Gdf9 mRNA level in postnatal day 14 (PD14) ovaries. (B, C) Western blotting results showing GDF9, SMAD3, and p-SMAD3 protein levels in PD14 ovaries. (D) p-SMAD3 staining (green) in PD14 ovaries. (E) GDF9 staining (red) in gene ontologies (GOs) from negative control (NC) and si-Matr3. n ≥68. (F) Quantification of the mean fluorescence intensity of GDF9 in oocytes. (G) Representative image of follicles and oocytes from NC, si-Matr3, si-Matr3+GDF9, GDF9. (H) Percentage of antral follicles and MII oocytes in (G). n≥3 per group. Scale bar: 40 μm in (D) and (E), 80 μm in (G). Data are represented as mean ± SD. ***p < 0.001, **p < 0.01, *p < 0.05, n.s., not significant.
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Figure 4—source data 1
PDF file containing original western blots for Figure 4B, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig4-data1-v1.zip
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Figure 4—source data 2
Original files for western blot analysis displayed in Figure 4B.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig4-data2-v1.zip
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Figure 4—source data 3
Source data for Figure 4A, C, F and H.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig4-data3-v1.xlsx
Figure 5 with 2 supplements
MATR3 promoted GDF9 by recruiting KDM3B.
(A) KDM3B staining (red) in gene ontology (GO) collected from negative control (NC) and si-Matr3. n ≥18. (B) Quantification of the mean fluorescence intensity of KDM3B (A) in oocytes. (C, D) Co-immunoprecipitation (co-IP) showing protein interactions between MATR3 and KDM3B in HEK293T cells. (E) Co-IP results after HEK293T cells were treated with pCDNA3.1-EGFP, pCDNA3.1-Matr3-EGFP (Matr3-FL-EGFP), pCDNA3.1-Matr3-ΔRRM2-EGFP (ΔRRM2-EGFP), or pCDNA3.1-Matr3-ΔpNLS4-EGFP (ΔpNLS4-EGFP) plasmid. (F) Live-cell imaging of oocytes after injecting Matr3-EGFP mRNA for 12 hr. (G) Live-cell imaging of oocytes after injecting EGFP, Matr3-FL-EGFP, ΔRRM2-EGFP, or ΔpNLS4-EGFP mRNA for 12 hr. (H) MATR3 (red) and GDF9 (green) staining in GO which had knocked down MATR3 protein before injected Matr3-FL, Matr3-ΔRRM2, or Matr3-ΔpNLS4 mRNA. n ≥36. (I) Quantification of the mean fluorescence intensity of GDF9 (H) in oocytes. Scale bar: 40 μm. Data are represented as mean ± SD. ***p<0.001, n.s., not significant.
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Figure 5—source data 1
PDF file containing original western blots for Figure 5C, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-data1-v1.zip
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Figure 5—source data 2
Original files for western blot analysis displayed in Figure 5C.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-data2-v1.zip
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Figure 5—source data 3
PDF file containing original western blots for Figure 5D, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-data3-v1.zip
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Figure 5—source data 4
Original files for western blot analysis displayed in Figure 5D.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-data4-v1.zip
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Figure 5—source data 5
Source data for Figure 5B, I.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-data5-v1.xlsx
Figure 5—figure supplement 1
The expression pattern of KDM3B in the ovaries of mice.
(A) Immunohistochemistry results showing KDM3B location in mice ovaries. (B) Immunofluorescence results showing the location of KDM3B in gene ontology (GO) and fully grown oocyte (FGO). (C) Western blotting results showing KDM3B protein levels in GO and FGO. Scale bar: 50 μm.
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Figure 5—figure supplement 1—source data 1
PDF file containing original western blots for Figure 5—figure supplement 1C, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-figsupp1-data1-v1.zip
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Figure 5—figure supplement 1—source data 2
Original files for western blot analysis displayed in Figure 5—figure supplement 1C.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-figsupp1-data2-v1.zip
Figure 5—figure supplement 2
Construction and localization detection of MATR3 truncated forms.
(A) Western blotting was used to detect the protein expression levels of the 293 cell line after being transfected with negative control (NC), MATR3, EGFP, and MATR3-EGFP plasmids for 48 hr, respectively. β-Actin is used as an internal reference to calibrate the protein loading level. (B) Immunofluorescence was used to detect the protein localization of the 293 cell line after transfection with MATR3-EGFP, ΔZnF1-EGFP, ΔZnF2-EGFP, ΔRRM1-EGFP, ΔRRM2-EGFP, and ΔpNL4-EGFP plasmids for 48 hr. (C) Immunofluorescence was used to detect the effect of the functional domains of MATR3 on its localization in living gene ontology (GO). Green: EGFP, MATR3 forming a fusion protein with EGFP; blue: Hoechst (nucleus). Scale bar: 10 μm in (B), 25 μm in (C).
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Figure 5—figure supplement 2—source data 1
PDF file containing original western blots for Figure 5—figure supplement 2A, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-figsupp2-data1-v1.zip
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Figure 5—figure supplement 2—source data 2
Original files for western blot analysis displayed in Figure 5—figure supplement 2A.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig5-figsupp2-data2-v1.zip
Figure 6 with 1 supplement
MATR3 is required for the oocyte-derived mushroom-like microvilli (OO-Mvi) in mouse oocyte by regulating Rdx mRNA level.
(A) p-ERM staining (green) showing the OO-Mvi in antral follicles (AFs) from Ctrl and cKO. (B) Quantification of the number of Oo-Mvi vesicles (n = 20). (C) Pie chart showing the proportion of the reads values of 10- to 12-day-old mice MATR3 low-input affinity cleavage enrichment sequencing (LACE-seq). (D) Table showing the base sequences to which MATR3 mainly binds. (E, G) Venn diagrams (E) and table (G) showing overlapping genes binding with MATR3, upregulated genes, and downregulated genes in single-cell RNA-sequencing (scRNA-seq). (F) Key gene ontology (GO) enrichment of all differentially expressed genes (DEGs) in LACE-seq. (H) IGV snapshot of MATR3 peaks distribution in 10- to 12-day-old mice ovaries. (I, G) RNA immunoprecipitation (RIP) showing the interaction between MATR3 and Rdx mRNA in 10- to 12-day-old mice ovaries. Western blotting showing the reliability of MATR3 antibody (I). Enrichment degrees of MATR3 on the Rdx mRNA, respectively (J). IgG served as the negative control and input (2%) served as the positive control. n=3. (K) RT-qPCR results showing Rdx mRNA level in postnatal day 14 (PD14) oocytes. Scale bar: 20 μm. Data are represented as mean ± SD. ***p<0.001.
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Figure 6—source data 1
PDF file containing original western blots and gels for Figure 6I, indicating the relevant bands and treatments.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig6-data1-v1.zip
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Figure 6—source data 2
Original files for western blot and gel analysis displayed in Figure 6I.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig6-data2-v1.zip
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Figure 6—source data 3
Source data for Figure 6 B, F, J and K.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig6-data3-v1.xlsx
Figure 6—figure supplement 1
Targets with high binding intensity and significant expression changes from the low-input affinity cleavage enrichment sequencing (LACE-seq) data.
RT-qPCR results showing Igf2bp2 and Ccnb1 mRNA level in postnatal day 14 (PD14) oocytes. Data are represented as mean ± SD. ***p<0.001.
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Figure 6—figure supplement 1—source data 1
Source data for Figure 6—figure supplement 1.
- https://cdn.elifesciences.org/articles/110703/elife-110703-fig6-figsupp1-data1-v1.xlsx
Figure 7
A proposed model: MATR3 regulates the synthesis and secretion of oocyte-secreted factors (OSFs), such as GDF9, thereby ensuring growing oocyte (GO) growth.
Under physiological conditions, MATR3 in the GOs promotes the transcription of a large number of genes required for cell growth. When promoting the transcription of the Gdf9 gene, MATR3 localizes in the nucleus through the pNLS4 domain to recruit KDM3B, thereby promoting the high-level expression of Gdf9. In ensuring the secretion of GDF9, MATR3 directly binds to Rdx mRNA to promote the growth of microvilli, which in turn lays the foundation for ensuring the secretion of factors, such as GDF9, and establishing the physical connection between oocytes and somatic cells.
Author response image 1
Ratio and survival rate of antral follicles after 6 days of culture.
n = 3. Data are represented as mean ± SD.
Author response image 2
Results of in vitro fertilization of oocytes.
2-cell: 2 days after fertilization; blastocyst: 4 days after fertilization. Data are represented as mean ± SD. ***p<0.001.
Author response image 3
Matr3 knockout results in the reduction of transcriptional activity.
(A) EU staining (green) in GO collected from Ctrl and cKO. n = 15. (B) Quantification of the mean fluorescence intensity of EU in oocytes. (C) H3K9me2 staining (red) in GO collected from Ctrl and cKO. n = 15. (D) Quantification of the mean fluorescence intensity of H3K9me2 in oocytes. Scale bar: 20 μm. Data are represented as mean ± S.D. ***P < 0.001.
Author response image 4
Matr3 knockdown impairs the structural integrity of oocyte OO-MVi.
(A) p-ERM staining (green) showing the OO-Mvi in oocyte from NC and si-Matr3. (B) Quantification of the number of Oo-Mvi vesicles (n = 6). Scale bar: 20 μm. Data are represented as mean ± SD. ***P < 0.001.
Author response image 5
Matr3 knockout has no effect on KDM3B localization.
KDM3B staining (red) in GO collected from Ctrl and cKO. n = 50. Scale bar: 40 μm.
Author response image 6
Loss of MATR3 in oocytes does not affect the number and proliferation of granulosa cells in primordial follicles.
(A) Immunohistochemistry results showing granulosa cells in PD7 ovaries from Ctrl and cKO. (B) Quantification of granulosa cell number in the largest cross-section of primary follicles. (C) Quantification of the proliferation rate of granulosa cells in the largest cross-section of primary follicles. n = 15. Data are represented as mean ± SD. n.s., not significant.
Additional files
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Supplementary file 1
gRNA and primer sequences for Matr3 knockout mouse construction and genotyping.
- https://cdn.elifesciences.org/articles/110703/elife-110703-supp1-v1.docx
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Supplementary file 2
RT-PCR primer sequences for target gene detection.
- https://cdn.elifesciences.org/articles/110703/elife-110703-supp2-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/110703/elife-110703-mdarchecklist1-v1.docx
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MATR3 is essential for oocyte growth and maturation quality through a dual molecular mechanism
eLife 15:RP110703.
https://doi.org/10.7554/eLife.110703.3
