Journal: medRxiv

Article Title: Physiological and molecular characterization of individuals carrying a diabetogenic mtDNA mutation establishes a mitochondrial basis for insulin resistance in humans

doi: 10.64898/2025.12.17.25342274

Figure Lengend Snippet: (A-D) Time course of venous and arterial plasma GDF15 levels during the hyperinsulinemic-euglycemic (HE) clamp. (B) GDF15 levels in venous plasma samples obtained before (Basal) and during the steady-state period of HE clamp (Insulin). (C) Time course of the venous-arterial difference in plasma GDF15 levels during the HE clamp. (D) Venous-arterial difference in plasma GDF15 before (Basal) and during the steady-state period of the HE clamp (Insulin). (E) Time course of venous and arterial plasma FGF21 levels during the HE clamp. n = 29 (15 in m.3243A>G, 14 in Controls). (F) FGF21 levels in venous plasma samples obtained before (Basal) and during the steady-state period of the HE clamp (Insulin). n = 29 (15 in m.3243A>G, 14 in Controls). (G) Time course of the venous-arterial difference in plasma FGF21 levels during the HE clamp. n = 29 (15 in m.3243A>G, 14 in Controls). (H) Venous-arterial difference in plasma FGF21 before (Basal) and during the steady-state period of the HE clamp (Insulin). n = 29 (15 in m.3243A>G, 14 in Controls). Linear mixed models were used to estimate within– and between-group differences [(B), (D), (F), and (H)]. Data are presented as observed individual values (with lines connecting individually matched participants) and estimated means ± 95% confidence limits, unless otherwise stated. For m.3243A>G carriers, individual datapoints are color-shaded to indicate muscle mtDNA heteroplasmy (light red = low, dark red = high). *Different from Basal ( P < 0.05). † Different from zero ( P < 0.05). n = 30 unless otherwise stated.

Article Snippet: Arterial and venous plasma samples were also analyzed for GDF15 and FGF21 using the automated immunoassay platform Ella (ProteinSimple, San Jose, California, USA) with Simple Plex assays for human GDF15 and FGF21 detection, according to the manufacturer’s instructions.

Techniques: Clinical Proteomics

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: GDF9 and BMP15 together, but not separately, induce AMH/Amh expression in vitro and in vivo. Effects of different concentrations of (A) GDF9 alone or (B) BMP15 alone on expression of AMH mRNA levels in KGN cells. Cotreatment of GDF9 and BMP15 in a dose-dependent manner significantly increases AMH/Amh mRNA levels compared with GDF9 or BMP15 alone in (C) KGN cells and (E) primary mGCs. Representative western blots with different concentrations of GDF9 and BMP15 treatment of 24 hours show increased AMH protein levels in (D) KGN cells and (F) mGCs. Serum AMH levels in 8-week-old mice (n = 5 animals per treatment) treated with vehicle (0 ng), with GDF9 or BMP15 alone (0.01 or 0.1 ng) or cotreated with GDF9 + BMP15 (0.001 ng) daily for 4 weeks. AMH/Amh mRNA data are displayed as mean ± SE (n = 3 experiments for both KGN cells and mGCs) and normalized to GAPDH/Gapdh levels. *P ≤ 0.05 vs control, 0 ng/mL; **P ≤ 0.01 vs control, GDF9 and BMP15 alone and ***P ≤ 0.01 vs control and GDF9 + BMP15 (0.5 and 1 ng/mL).

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, In Vitro, In Vivo, Western Blot, Control

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: GDF9 and BMP15 induce AMH expression through Smad2/3 and PI3K/Akt signaling in KGN cells. Relative expression of (A) AMH mRNA levels and (B) AMH protein levels in GDF9 + BMP15 (2.5 ng/mL) stimulated KGN cells pretreated with PKA inhibitor H89 (10 μM), PI3K inhibitor LY294002 (10 μM), MAPK inhibitor U0126 (10 μM), Smad2/3 inhibitor SB431542 (10 μM), and Smad 1/5/8 inhibitor LDN193189 (100 nM). (C, D) GDF9 + BMP15 induces (C) Smad2/3 and (D) Akt activation in KGN cells. (E) Inhibition of Smad2/3 pathway with SB431542 (10 μM) has no effect on GDF9 + BMP15–induced Akt phosphorylation. (F, G) PI3K/Akt pathway alone is not sufficient to induce AMH expression. Insulin stimulates Akt but not (E) Smad2/3 phosphorylation and has no effect on the (F) expression of AMH mRNA levels in KGN cells. AMH mRNA data are displayed as mean ± SE (n = 3 experiments) and normalized to GAPDH levels. *P ≤ 0.001 vs media. LDN, LDN193189; LY, LY294002; SB, SB431542.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Activation Assay, Inhibition, Phospho-proteomics

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: GDF9 and BMP15 induce AMH expression through Smad2/3 and PI3K/Akt signaling in mGC cultures. Relative expression of (A) Amh mRNA levels and (B) AMH protein levels in GDF9 + BMP15 (2.5 ng/mL) stimulated mGCs pretreated with PKA inhibitor H89 (10 μM), PI3K inhibitor LY294002 (10 μM), MAPK inhibitor U0126 (10 μM), Smad2/3 inhibitor SB431542 (10 μM), and Smad 1/5/8 inhibitor LDN193189 (100 nM). (C, D) GDF9 + BMP15 induce (C) Smad2/3 and (D) Akt activation. AMH mRNA data are displayed as mean ± SE (n = 3 experiments) and normalized to Gapdh levels. *P ≤ 0.001 vs media. LDN, LDN193189; LY, LY294002; SB, SB431542.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Activation Assay

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: GDF9 + BMP15 through the PI3K/Akt pathway phosphorylates p300 that is essential for AMH expression. GDF9 + BMP15 (2.5 ng/mL) treatment phosphorylates p300 at ser1834 that is blocked by the PI3K inhibitor LY294002 (10 μM) in (A) KGN cells and (D) mGCs. siRNA-mediated knockdown of p300 in (B) KGN cells and (E) mGCs (inset) inhibits (C, E) GDF9 + BMP15–mediated increase in AMH/Amh mRNA levels. Nontargeting siRNA (Nsp) pool is used as control. AMH/Amh mRNA data are displayed as mean ± SE (n = 3 experiments for both KGN cells and mGCs) and normalized to GAPDH/Gapdh levels. *P ≤ 0.001 vs Nsp and media. LY, LY294002.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Knockdown, Control

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: GDF9 + BMP15 treatment increases p300 binding and H3K27 acetylation mark on the AMH promoter. (A) Anti-p300 and anti-H3K27ac ChIP assay in KGN cells showing the amount of p300 binding and level of H3K27ac on the AMH promoter (P1 region) following GDF9 + BMP15 (2.5 ng/mL) treatment in the presence or absence of the PI3K inhibitor LY294002 (10 μM) and Smad2/3 inhibitor SB431542 (10 μM). Values represent percentage input (mean ± SE, n = 3 experiments). *P ≤ 0.01 vs media. (B, C) Smad2/3 and p300 form a complex in the nucleus following GDF9 + BMP15 treatment. Smad2/3 or p300 was precipitated from nuclear extracts of GDF9 + BMP15 (2.5 ng/mL)–treated (B) KGN cells and (C) mGCs in the absence or presence of PI3K inhibitor LY294002 (10 μM) or Smad2/3 inhibitor SB431542 (10 μM), followed by immunoblotting (IB) (n = 3 with identical results for both KGN cells and mGCs). LY, LY294002; SB, SB431542.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Binding Assay, Western Blot

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: FSH inhibits GDF9 + BMP15–induced AMH expression. FSH treatment blocks GDF9 + BMP15–induced AMH/Amh expression in (A) KGN cells and (B) mGC culture. (B and C, insets) Representative western blots showing FSH treatment decreases GDF9 + BMP15–induced increase in AMH protein levels in KGN cells and mGCs. Pretreatment of (B) KGN cells and (D) mGCs with the PKA inhibitor H89 (10 μM) and SF1 inhibitor SID7969543 (1 μM) abrogates the inhibitory effects of FSH (5 ng/mL) on GDF9 + BMP15 (2.5 ng/mL)–induced AMH expression. AMH/Amh mRNA data are displayed as mean ± SE (n = 3 experiments for both KGN cells and mGCs) and normalized to GAPDH/Gapdh levels. *P ≤ 0.01 vs GDF9 + BMP15, FSH (0 ng/mL); **P ≤ 0.05 vs GDF9 + BMP15, FSH (0, 1, 10, and 50 ng/mL); ***P ≤ 0.05 vs GDF9 + BMP15, FSH (0, 1, and 5 ng/mL). SID, SID7969543.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Western Blot

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: Inhibition of GDF9 + BMP15–induced AMH expression by FSH is mediated through HDAC enzymes. (A) Pretreatment of KGN cells with trichostatin A (TSA; 100 nM), a broad-spectrum HDAC inhibitor, reverses the inhibitory effects of FSH (5 ng/mL) on GDF9 + BMP15 (2.5 ng/mL)–induced AMH expression. (B) KGN cells were treated with siRNAs (100 nM) against different HDAC isoforms, and its effect on the FSH-mediated decrease on GDF9 + BMP15–induced AMH expression was determined. siRNA-mediated knockdown of HDAC2 rescues AMH/Amh mRNA levels in KGN cells and primary mGCs (C) stimulated with GDF9 + BMP15 (2.5 ng/mL) and treated with FSH (5 ng/mL). Nonspecific siRNA (Nsp) pool is used as control. AMH/Amh mRNA data are displayed as mean ± SE (n = 3 experiments) and normalized to GAPDH/Gapdh levels. *P ≤ 0.001 vs control.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Inhibition, Expressing, Knockdown, Control

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: FSH treatment decreases the H3K27ac mark through increased HDAC2 binding on the AMH promoter. Anti-H3K27ac and anti-HDAC2 ChIP assay in GDF9 + BMP15 (2.5 ng/mL) stimulated KGN cells or siRNA-mediated GIOT1 knocked down KGN cells, showing the level of H3K27ac and the amount of HDAC2 binding on the AMH promoter (P1 region) following FSH treatment. Values represent percentage input (mean ± SE, n = 3 experiments). *P ≤ 0.001 vs media.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Binding Assay

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: FSH inhibits AMH expression through inducing the expression of GIOT1. FSH (5 ng/mL) treatment induces the expression of GIOT1/Giot1 mRNA levels that is blocked by the PKA inhibitor H89 (10 μM) and the SF1 inhibitor SID7969543 (1 μM) in (A) KGN cells and (D) mGCs. siRNA-mediated knockdown of GIOT1/Giot1 in (B) KGN cells and (E) mGCs inhibits the (C, F) GDF9 + BMP15–mediated increase in AMH/Amh mRNA levels. Nonspecific siRNA (Nsp) pool is used as control. GIOT1/Giot1 and AMH/Amh mRNA data are displayed as mean ± SE (n = 3 experiments for both KGN and mGCs) and normalized to GAPDH/Gapdh levels. *P ≤ 0.001 vs Nsp and media/control. SID, SID7969543.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Knockdown, Control

Journal: Endocrinology

Article Title: Oocyte-Derived Factors (GDF9 and BMP15) and FSH Regulate AMH Expression Via Modulation of H3K27AC in Granulosa Cells

doi: 10.1210/en.2018-00609

Figure Lengend Snippet: (A) Expression of Amh mRNA in ovaries isolated from WT, Fshβ-null mice (Fshβ−/−), and Fshβ-null mice expressing human WT FSH (Fshβ−/−hFSHβWT) mice. Amh mRNA data are displayed as mean ± SE (n = 3 animals per genotype) and normalized to Gapdh levels. *P ≤ 0.001 vs WT. (B) Proposed model for regulation of AMH expression by GDF9 + BMP15 and FSH. GDF9 + BMP15 phosphorylates p300 at ser1834, resulting in its transactivation. GDF9 + BMP15 also activate Smad2/3, and the latter interacts with p300 and recruits it on the AMH promoter, causing acetylation of lysine 27 on histone 3 (H3K27ac), resulting in AMH gene expression. FSH, through the PKA/SF-1 pathway, induces the expression of GIOT1 that recruits HDAC2 to the AMH promoter, causing deacetylation of GDF9 + BMP15–induced H3K27ac, thereby inhibiting AMH expression.

Article Snippet: For in vivo studies, 8-week-old mice were injected intraperitoneally with different concentrations of recombinant human GDF9 (R&D Systems) and/or BMP15 (R&D Systems) once daily for 4 weeks (n = 5 mice per treatment).

Techniques: Expressing, Isolation, Gene Expression

Journal: bioRxiv

Article Title: Lac-Phe mediates the anti-obesity effect of metformin

doi: 10.1101/2023.11.02.565321

Figure Lengend Snippet: (A) 300 mg/kg metformin treatment increases Lac-Phe levels in WT (n = 6), but not in CNDP2-KO (n = 7) mice. (B) Chronic metformin treatment (300 mg/kg daily) suppresses food intake of WT, but not CNDP2-KO mice. (C) Chronic metformin treatment (300 mg/kg daily) suppresses body weight of WT, but not CNDP2-KO mice. (D) 300 mg/kg metformin treatment increases circulating GDF15 levels in WT and CNDP2-KO mice. (E-F) 4 nmol subcutaneous GDF15 injection reduces food intake (E) and body weight (F) in WT and CNDP2-KO mice. Values adjusted by effects with vehicle treatment. (G) 300 mg/kg metformin treatment increases circulating GLP-1 levels in WT and CNDP2-KO mice. (H-I) 10 nmol subcutaneous Semaglutide injection reduces food intake (H) and body weight (I) in WT and CNDP2-KO mice. Values adjusted by effects with vehicle treatment. For (B) and (C), N = 6 for KO-saline, N = 7 for other groups. N = 4 – 5 in (D) and (G). N = 5 – 6 in (E), (F), (H), and (I). P values in (A) were calculated using multiple paired t tests with Holm-Šídák corrections; Welch t tests were used in (D-I).

Article Snippet: Recombinant GDF15 (957-GD) was purchased from R&D systems.

Techniques: Injection, Saline

Journal: MethodsX

Article Title: A method of quantitative chemiluminescence immunoassay for the concentration of Growth differentiation factor-15

doi: 10.1016/j.mex.2024.102572

Figure Lengend Snippet: A, Correlation between GDF-15 levels in 254 serum samples were measured by CLIA vs. ELISA. Correlation coefficient and P-value were calculated using the Spearman's rank correlation. B, Differences in GDF-15 concentrations between CLIA and ELISA. C, Median and interquartile range (IQR) of GDF-15 concentrations for CLIA and ELISA.

Article Snippet: To analyze the effectiveness of this chemiluminescence immunoassay, we performed methodological comparisons with commercially available ELISA kits (Human GDF-15 Quantikine QuicKit ELISA, Catalog: QK957, R&D systems).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: MethodsX

Article Title: A method of quantitative chemiluminescence immunoassay for the concentration of Growth differentiation factor-15

doi: 10.1016/j.mex.2024.102572

Figure Lengend Snippet:

Article Snippet: To analyze the effectiveness of this chemiluminescence immunoassay, we performed methodological comparisons with commercially available ELISA kits (Human GDF-15 Quantikine QuicKit ELISA, Catalog: QK957, R&D systems).

Techniques: Chemiluminescence Immunoassay, Enzyme-linked Immunosorbent Assay

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 1. Transzonal projections (TZP) density of COCs during IVM with BMP15 (A) and/or GDF9 (B). Phalloidin-FITC fluorescence intensity (mean ± S.E.M.) in the zona pellucida area of COCs after 0h, 6h, 12h and 24h of IVM. A total of 20–30 COCs per group were assessed in 3 replicates. (A) Values with different letters differ significantly (p < 0.05). (B) Different letters between groups in the same timepoint (a,b) and different symbols between timepoints in the same group (α,β,γ for control and GDF9 groups; and σ,τ for GDF9+BMP15 group) indicate significant differences (p < 0.05). (C) Representative confocal images of phalloidin-FITC staining. TZP are observed as thin filaments going through the zona pellucida from the cumulus cells to the oocyte. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence, Control, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 2. Mitochondrial activity of MII-oocytes after IVM with BMP15 and/or GDF9. (A) Relative fluorescence intensity (mean ± S.E.M.) of Mitotracker stained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 20–27 oocytes per group were assessed in 3 replicates. (B) Representative images. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activity Assay, Fluorescence, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 3. ROS and GSH levels after IVM with BMP15 and/or GDF9. Relative fluorescence intensity (mean ± S.E.M.) of (A) ROS and (B) GSH. A total of 24–31 oocytes per group were assessed in 3 replicates. (C) Representative images. Scale bar: 200 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 4. EGFR expression of MII-oocytes after IVM with BMP15 (A) and/or GDF9 (B). Relative fluorescence intensity (mean ± S.E.M.) of EGFR immunostained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 26–40 oocytes per group were assessed in 4 replicates. (C) Representative images. Scale bar: 30 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 5. EGFR expression of cumulus cells after IVM with BMP15 (A) and/or GDF9 (B). Relative EGFR expression (mean ± S.E.M.). Columns with different letters differ significantly (p < 0.05). Four independent blots were used for relative quantification. (C) Western blot analyses of EGFR expression. EGFR from 70 to 80 COCs/ replicate was quantified by western blotting and standardized to vinculin protein levels.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Quantitative Proteomics, Western Blot

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 6. Cumulus expansion index after IVM with BMP15 (A) and/or GDF9 (B). Cumulus expansion of each COC was scored on a 0 to 4 scale and cumulus expansion index (mean ± S.E.M) was calculated as the arithmetic mean value scored of all COCs evaluated per replica. Columns with different letters differ significantly (p < 0.05). A total of 367–441(A) and 278–323 (B) oocytes per group were assessed in 5 replicates.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques:

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 7. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after parthenogenic activation (PA). Cell number (mean ± S.E. M.) of the inner cell mass (ICM) and the trophectoderm (TE) of 8-days blastocysts. Columns with different letters differ significantly (p < 0.05). A total of 15–29 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activation Assay

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 8. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after IVF. Cell number (mean ± S.E.M.) of 8-day blastocysts in the inner cell mass (ICM) and the trophectoderm (TE). Columns with different letters differ significantly (p < 0.05). A total of 14–22 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques:

Journal: Frontiers in endocrinology

Article Title: Acute endurance exercise modulates growth differentiation factor 11 in cerebrospinal fluid of healthy young adults.

doi: 10.3389/fendo.2023.1137048

Figure Lengend Snippet: FIGURE 1 An acute bout of endurance exercise (90 min run) modified the levels of GDF11 in cerebrospinal fluid but not in circulation of healthy trained volunteers. GDF11 levels were assessed by immunoblotting, using three different primary antibodies: (A) GDF11-specific N-terminus antibody (CSF, n=12/12, B/R, p=0.046); (B) GDF11-specific C-terminus antibody (CSF, n=13/13, B/R, p=0.008); (C) GDF11/8 antibody (CSF, n=12/12, B/R, p=0.065); (D) GDF11-specific N-terminus antibody (serum, n= 14/14/14, B/0/R); (E) GDF11-C-terminus antibody (plasma, n=13/13/13, B/0/R); (F) GDF11- specific N-terminus antibody (plasma, n=14/14/13, B/0/R); Due to the lack of space, molecular weight ladders are not visible in Figures 1D–F and therefore, the molecular weights of GDF11 are in this case approximate, based on our validation studies of antibodies. B, Basal, before run; R, after run; A.U., signal intensity; GDF, Growth Differentiation Factor. Immunoblotting of all samples was performed under identical conditions. Statistical differences were analysed using paired Student’s t-test. *p<0.05, **p<0.01, #p<0.1 (A–C).

Article Snippet: To corroborate the findings, GDF11 C-terminus specific monoclonal antibody was used (recombinant human GDF11 aa299-407, 1:1000, R&D systems, MAB19581, USA) (sample set B, n=13; overlap between A & B sample sets: n=7, Supplementary Figure 1B).

Techniques: Western Blot, Clinical Proteomics, Molecular Weight, Biomarker Discovery

Journal: Frontiers in endocrinology

Article Title: Acute endurance exercise modulates growth differentiation factor 11 in cerebrospinal fluid of healthy young adults.

doi: 10.3389/fendo.2023.1137048

Figure Lengend Snippet: FIGURE 2 Running-induced decrease of blood-brain barrier permeability (CSF/serum albumin ratio). n=17. (albumin values were not measured in CSF of one individual, and two individuals had CSF available only after the run) (A); its association with CSF/serum GDF11 ratio (B); and CSF/serum GDF11/8 ratio (C). CSF, Cerebrospinal Fluid; GDF, Growth Differentiation Factor. ***p<0.001; Open circles - before run; Full circles - after run.

Article Snippet: To corroborate the findings, GDF11 C-terminus specific monoclonal antibody was used (recombinant human GDF11 aa299-407, 1:1000, R&D systems, MAB19581, USA) (sample set B, n=13; overlap between A & B sample sets: n=7, Supplementary Figure 1B).

Techniques: Permeability

Journal: Frontiers in endocrinology

Article Title: Acute endurance exercise modulates growth differentiation factor 11 in cerebrospinal fluid of healthy young adults.

doi: 10.3389/fendo.2023.1137048

Figure Lengend Snippet: FIGURE 3 Heat map depicting associations between levels of GDF11 and clinical parameters, as well as their running-induced changes. Data were analyzed using GDF11-specific N-terminus antibody (paired serum samples n=14; paired CSF samples n=12, unpaired CSF samples available only after run n=2). BMI, body mass index; CSF, cerebrospinal fluid; GDF, Growth Differentiation Factor; HRmax, maximal heart rate; S-CK, serum creatinkinase; R, correlation coefficient; VO2max, maximal aerobic capacity. Coefficients of significant correlations (p<0.05) or trends (p<0.1) are presented in numbers.

Article Snippet: To corroborate the findings, GDF11 C-terminus specific monoclonal antibody was used (recombinant human GDF11 aa299-407, 1:1000, R&D systems, MAB19581, USA) (sample set B, n=13; overlap between A & B sample sets: n=7, Supplementary Figure 1B).

Techniques:

Journal: Endocrinology

Article Title: TGFβ Superfamily Members Mediate Androgen Deprivation Therapy-Induced Obese Frailty in Male Mice.

doi: 10.1210/en.2016-1580

Figure Lengend Snippet: Figure 7. Castration induces myostatin protein levels in skeletal muscle. A, Representative immunoblots of myostatin and -actin (reprobing of the myostatin blot) expression in GAS muscle. B, Quantification of mean myostatin levels in GAS muscles from 4 mice at each time, assessed by 3 independent measurements. C, Representative immunoblot of myostatin and - actin (reprobing of the myostatin blot) expression level in TRI muscle. Lanes of immunoblots marked (C) contain identical control sample for interblot comparison. D, Quantification of mean myostatin levels in TRI muscles from 4 mice at each time, assessed by 3 independent measurements. The values shown in B and D are relative to the sham-castrated (0-wk castrate) animals. Sham-castrated (blue) and castrated groups (red) were compared by one-way ANOVA with Dunnett’s testing. Bars are SEM; *, P .05; **, P .01; ***, P .001 vs sham-castrated group.

Article Snippet: Target Antigen Sequence (if Known) Name of Antibody Manufacturer, Catalog Number, and/or Name of Individual Providing the Antibody Species Raised in; Monoclonal or Polyclonal Dilution Used Myostatin Mouse GDF8 Asn25Ser376 MAb (clone 84214) R&D Systems, MAB788 Monoclonal; rat IgG2B 1:1000 Phospho-Smad2 Smad3S423/5 MAb (clone EP823Y) Abcam, 52903 Monoclonal; rabbit 1:2000 Smad2 Amino terminus hSmad2 L16D3 Cell Signaling, 3103 Monoclonal; mouse 1:2000 Eukaryotic EF2 Complete protein G270 Nastiuk (Ref. 63) Rabbit 1:2000 -Actin -Actin aa 1–14 AC15 Sigma, A5441 Monoclonal; mouse 1:5000 Rat IgG (H L) Rabbit antirat unconjugated Pierce, 31218 Rabbit 1:20 000 Rabbit IgG (H L) Goat antirabbit HRP- conjugated secondary Pierce, 31460 Goat 1:20 000 Mouse IgG (H L) Goat antimouse HRPconjugated secondary Pierce, 31430 Goat 1:20 000 D ow nloaded from https://academ ic.oup.com /endo/article/157/11/4461/2758425 by G ITAM (D eem ed to be U niversity) user on 01 January 2025 tion are an acutely sensitive biological measure of circulating androgen elimination (34, 35).

Techniques: Western Blot, Expressing, Muscles, Control, Comparison