Journal: Cell Communication and Signaling : CCS

Article Title: GDF9 His209GlnfsTer6/S428T and GDF9 Q321X/S428T bi-allelic variants caused female subfertility with defective follicle enlargement

doi: 10.1186/s12964-024-01616-8

Figure Lengend Snippet: Effects of GDF9 variants on gene expression and GDF9-BMP15 interactions. A Western blots of mature GDF9 proteins expressed in follicular fluid collected from the first and second retrieval cycle of individual P1 and age-matched controls ( n =3). B ELISA of GDF9 in follicular fluid collected from the first retrieval cycle of individual P1 and age-matched controls ( n =6). C Western blots of GDF9 expression in HEK293T cells transfected with human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative mature-GDF9 expression in culture medium (CM) (3 independent experiments). The amount of concentrated culture medium used for the detection was 35μl per sample per assay. lnfsTer6, His209GlnfsTer6. * P < 0.05. D Western blots of GDF9 precursors and mature GDF9 over time during in vitro cleavage assay (3 independent experiments). Total GDF9=GDF9 precursor+mature GDF9. E Western blots of BMP15 expression in HEK293T cells co-transfected with human BMP15 and human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative BMP15 precursor expression in CM (3 independent experiments). lnfsTer6, His209GlnfsTer6. * P < 0.05. F Co-IP assay of the binding of GDF9 proteins to BMP15 protein. lnfsTer6, His209GlnfsTer6. * P < 0.05. G Western blots of p-smad2 and smad2 expression in human primary luteinized granulosa cells treated with purified recombinant pro-GDF9 WT -BMP15 and pro-GDF9 S428T -BMP15 protein respectively (3 independent experiments)

Article Snippet: GDF9 was measured in follicular fluid collected at the first oocyte retrieval from individual P1 and in follicular fluid from other six control women using an ELISA kit (CSB-E12925h, CUSABIO, China).

Techniques: Gene Expression, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, In Vitro, Cleavage Assay, Co-Immunoprecipitation Assay, Binding Assay, Purification, Recombinant

Journal: Heliyon

Article Title: Comparing GDF9 in mature follicles and clinical outcomes across different PCOS phenotype

doi: 10.1016/j.heliyon.2024.e29879

Figure Lengend Snippet: Distribution of GDF9 in FF from the three PCOS phenotypes and control group. Data are presented as mean ± SD. d P < 0.01, compared with Control Group; a P < 0.01, compared with Control Group; b P < 0.05, compared with PCOS A; c P < 0.05, compared with PCOS B.

Article Snippet: The FF was diluted 1:5 in phosphase-buffered saline (PBS) and measured using a human GDF9 ELISA Kit (Elabscience, Wuhan, China) according to the manufacture's instructions.

Techniques: Control

Journal: Heliyon

Article Title: Comparing GDF9 in mature follicles and clinical outcomes across different PCOS phenotype

doi: 10.1016/j.heliyon.2024.e29879

Figure Lengend Snippet: Expression of GDF9 in the CCs. (a) Immunohistochemical staining of GDF9 expressed in the control group, phenotype A, phenotype B and phenotype D. (Original magnification: × 400, bars, 50 μm). (b) Mean density of GDF9 in CCs of the three PCOS phenotypes and control group, a P < 0.01, compared with Control Group; b P < 0.01, compared with PCOS D; c P < 0.01, compared with Control Group; d P < 0.01, compared with PCOS D.

Article Snippet: The FF was diluted 1:5 in phosphase-buffered saline (PBS) and measured using a human GDF9 ELISA Kit (Elabscience, Wuhan, China) according to the manufacture's instructions.

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: Heliyon

Article Title: Comparing GDF9 in mature follicles and clinical outcomes across different PCOS phenotype

doi: 10.1016/j.heliyon.2024.e29879

Figure Lengend Snippet: Analysis of multiple factors affecting blastocyst formation and clinical pregnancy rate in the PCOS phenotypes.

Article Snippet: The FF was diluted 1:5 in phosphase-buffered saline (PBS) and measured using a human GDF9 ELISA Kit (Elabscience, Wuhan, China) according to the manufacture's instructions.

Techniques: Control

Journal: PLoS ONE

Article Title: Effect of Anti-Mullerian Hormone in Culture Medium on Quality of Mouse Oocytes Matured In Vitro

doi: 10.1371/journal.pone.0099393

Figure Lengend Snippet: Real-time PCR primer sequences.

Article Snippet: The procedure was performed with a GDF9 ELISA Kit (SC-12244, Santa Cruz, USA) and a BMP15 ELISA Kit (E03B0394, Blue Gene, China) following the manufacturer's instructions.

Techniques: Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Effect of Anti-Mullerian Hormone in Culture Medium on Quality of Mouse Oocytes Matured In Vitro

doi: 10.1371/journal.pone.0099393

Figure Lengend Snippet: GDF9 and BMP15 mRNA in the oocytes were measured by RT-PCR, and GDF9 and BMP15 proteins in IVM-medium were measured by ELISA, respectively. Oocytes were harvested at 16–18 h after IVM. *Indicates significant differences (p<0.05) compared to the control group. Data were from 3 replicates.

Article Snippet: The procedure was performed with a GDF9 ELISA Kit (SC-12244, Santa Cruz, USA) and a BMP15 ELISA Kit (E03B0394, Blue Gene, China) following the manufacturer's instructions.

Techniques: Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: PLoS ONE

Article Title: Effect of Anti-Mullerian Hormone in Culture Medium on Quality of Mouse Oocytes Matured In Vitro

doi: 10.1371/journal.pone.0099393

Figure Lengend Snippet: GDF9 and BMP15 mRNA expression in the oocytes were measured by RT-PCR, and GDF9, and BMP15 proteins in IVM-medium were measured by ELISA, respectively. Oocytes and IVM-medium supernatants were collected at 6, 12 and 18 h respectively during IVM. *Indicates significant differences (p<0.05) compared to the control group. Data were from 3 replicates.

Article Snippet: The procedure was performed with a GDF9 ELISA Kit (SC-12244, Santa Cruz, USA) and a BMP15 ELISA Kit (E03B0394, Blue Gene, China) following the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: PLoS ONE

Article Title: Effect of Anti-Mullerian Hormone in Culture Medium on Quality of Mouse Oocytes Matured In Vitro

doi: 10.1371/journal.pone.0099393

Figure Lengend Snippet: Cumulus cells and oocytes were harvested from COCs infected by AdH1-SiRNA/NS or AdH1-SiRNA/AMH during IVM, respectively. (A) Shows AMH mRNA expression in cumulus cells infected by AdH1-SiRNA/NS or AdH1-SiRNA/AMH; (B) Shows GDF9 and BMP15 mRNA expressions in oocytes from COCs infected by AdH1-SiRNA/NS or AdH1-SiRNA/AMH, respectively. *Indicates significant differences (p<0.05) compared to the control group. Data were from 3 replicates.

Article Snippet: The procedure was performed with a GDF9 ELISA Kit (SC-12244, Santa Cruz, USA) and a BMP15 ELISA Kit (E03B0394, Blue Gene, China) following the manufacturer's instructions.

Techniques: Infection, Expressing, Control