Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Relative mRNA expressions of Bmp15, Gdf9, and C-KIT in the ovaries of mice aged from 10 to 40 weeks. Levels of the mRNAs of Bmp15, Gdf9, and C-KIT were normalized to the amount of GAPDH per sample. Data are presented as mean ± standard deviation.*p < 0.05 (vs. 10 weeks old).

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Immunohistochemistry of BMP15, GDF9, and C-KIT in the mouse ovaries according to age. (a, e, i) 10 weeks, (b, f, j) 20 weeks, (c, g, k) 30 weeks, and (d, h, l) 40 weeks of mice. Immunostainings with anti-BMP15 (a–d) and cv-Kit (i–l) antibodies were usually localized in oocyte and cumulus cells, whereas immunostaining with anti-GDF antibody (e–h) was detected just in the oocytes (brown). (m) Quantitative results of immunohistochemistry analysis. Five ovaries were used per age group and a total of 10 follicles per ovary were counted for analysis. The images are representative at a final magnification of ×100. Scale bar indicates 50 µm. Black arrowhead, white arrowhead, black arrow, and red arrow indicate oocytes of primary, secondary, antral, and graffian follicles, respectively. *p < 0.05 (vs. 10 weeks old). (A color version of this figure is available in the online journal.)

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Immunohistochemistry, Immunostaining

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Protein expressions of BMP15, GDF9, and C-KIT in the ovaries of mice aged 10 to 40 weeks. Protein expression levels of BMP15, GDF9, and C-KIT were normalized to the amount of actin per sample. Relative protein densities were quantified using NIH-Image J software (version 1.35d). Data are presented as mean ± standard deviation. Results are representative of at least three independent experiments. *p < 0.05 (vs. 10 weeks old).

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Expressing, Software, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Expressions changes of Bmp15, Gdf9 and C-KIT in transcriptome analysis using RNA-Seq for ovaries of mice aged 6 weeks and 48 weeks. (a) Heat-map results, (b) quantification of expression level of mRNA. Expressions of BMP15, GDF9, and C-KIT were significantly decreased in old mice compared with young mice.

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: RNA Sequencing, Expressing

Journal: Reproduction (Cambridge, England)

Article Title: REGULATION OF AMH BY OOCYTE SPECIFIC GROWTH FACTORS IN HUMAN PRIMARY CUMULUS CELLS

doi: 10.1530/REP-17-0421

Figure Lengend Snippet: Cumulus cells were treated for 48 hours with vehicle (C), GDF9 (2.5, 10ng/ml), BMP15 (2.5, 10ng/ml), or the combination of GDF9 and BMP15 (0.1, 0.6, 2.5, 5, or 10ng/ml of each). Amh mRNA levels were determined by qPCR and expressed as relative to Rpl19. Columns represent the mean ± SEM, Columns with different letters differ significantly a–b P<0.05, a–c P<.001 vs controls, one-way ANOVA, Bonferroni test, n=15.

Article Snippet: Cumulus cells were treated with human recombinant GDF9 (R&D Systems), BMP15 (R&D Systems), and/or FSH (Serono) with or without specific inhibitors of intracellular signaling; H89, GF109203X (EMD Biosciences-Calbiochem), MK2206, LDN-193189 (Selleck Chemicals), U0126 (Millipore), or SB431542 (Tocris).

Techniques:

Journal: Reproduction (Cambridge, England)

Article Title: REGULATION OF AMH BY OOCYTE SPECIFIC GROWTH FACTORS IN HUMAN PRIMARY CUMULUS CELLS

doi: 10.1530/REP-17-0421

Figure Lengend Snippet: A) Cumulus cells from three different patients (Patient 1, 2, and 3) were treated with 5 ng/ml of both GDF9 and BMP15 (GB) for 48 hours. AMH β-actin (ACTB) protein levels were determined by Western blotting. On the left, the average (± SEM) of the relative optical density units of AMH to ACTB is shown (**P< 0.001, t-test, n=3). B) Cumulus cells from Patient 4 were treated with 2.5, 5, or 20 ng/ml of both GDF9 and BMP15. AMH protein levels were determined as in A. Expression of ACTB is shown as a loading control.

Article Snippet: Cumulus cells were treated with human recombinant GDF9 (R&D Systems), BMP15 (R&D Systems), and/or FSH (Serono) with or without specific inhibitors of intracellular signaling; H89, GF109203X (EMD Biosciences-Calbiochem), MK2206, LDN-193189 (Selleck Chemicals), U0126 (Millipore), or SB431542 (Tocris).

Techniques: Western Blot, Expressing, Control

Journal: Reproduction (Cambridge, England)

Article Title: REGULATION OF AMH BY OOCYTE SPECIFIC GROWTH FACTORS IN HUMAN PRIMARY CUMULUS CELLS

doi: 10.1530/REP-17-0421

Figure Lengend Snippet: Cumulus cells were treated for 48 hours with GDF9 and BMP15 (G+B; 5 ng/ml of each) in the presence or absence of FSH (50 ng/ml). Amh mRNA levels were determined by qPCR and expressed relative to Rpl19. Columns with different letters differ significantly a–b P<0.001, a–c P<0.05, b–c P<0.01, one-way ANOVA, Bonferroni test, n=11.

Article Snippet: Cumulus cells were treated with human recombinant GDF9 (R&D Systems), BMP15 (R&D Systems), and/or FSH (Serono) with or without specific inhibitors of intracellular signaling; H89, GF109203X (EMD Biosciences-Calbiochem), MK2206, LDN-193189 (Selleck Chemicals), U0126 (Millipore), or SB431542 (Tocris).

Techniques:

Journal: Reproduction (Cambridge, England)

Article Title: REGULATION OF AMH BY OOCYTE SPECIFIC GROWTH FACTORS IN HUMAN PRIMARY CUMULUS CELLS

doi: 10.1530/REP-17-0421

Figure Lengend Snippet: Cumulus cells were treated for 1 hour with the following specific SB431542 (SMAD2/3; 10 μM), or LDN-193189 (SMAD1/5/8; 100 nM) followed by a 48-hour co-treatment with GDF9 and BMP15 (5 ng/ml of each). Amh mRNA levels were determined 48 h after by qPCR and expressed relative to Rpl19. Columns with different letters differ significantly a–b P<0.001, one-way ANOVA, Bonferroni test, n=9.

Article Snippet: Cumulus cells were treated with human recombinant GDF9 (R&D Systems), BMP15 (R&D Systems), and/or FSH (Serono) with or without specific inhibitors of intracellular signaling; H89, GF109203X (EMD Biosciences-Calbiochem), MK2206, LDN-193189 (Selleck Chemicals), U0126 (Millipore), or SB431542 (Tocris).

Techniques: