Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: Msx2 is a downstream target of BMP2 signaling in the uterus during decidualization. (A) The primary cultures of mouse endometrial stromal cells (MESCs) were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points, as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of Msx2. The relative levels of gene expression were determined by setting the expression level of the GFP-treated sample at 24 h to 1.0 (n = 3). Rplp0, encoding a ribosomal protein, was used to normalize the level of RNA. *P < 0.05. (B) The nucleotide positions of the SBEs on the Msx2 promoter were analyzed by ChIP. (C) Mouse stromal cells were treated with E + P or E, P, and BMP2 (E + P + BMP2) for 90 min. ChIP, using the Smad4 antibody, was performed, as described in “Materials and Methods.” Chromatin enrichment was quantified by real-time PCR using primers flanking the potential SBE in the Msx2 promoter and also a negative control region in the ORF of Msx2. Enrichments were normalized to 1% of input DNA. The experiment was repeated twice, and representative data are shown.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression, Negative Control

Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: MSX1 and MSX2 mediate BMP2-induced HESC decidualization. The primary cultures of HESCs were established as described in “Materials and Methods.” The cells were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of MSX1 and MSX2. The relative levels of gene expression were determined by setting the expression level on day 0 of the GFP-treated sample at 1.0. RPLP0, encoding a ribosomal protein, was used to normalize the level of RNA. Data were collected from three independent clinical samples, which were subjected to the same experimental conditions. *P < 0.05; **P < 0.005.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Immunity

Article Title: Stellate Cells, Hepatocytes, and Endothelial Cells Imprint the Kupffer Cell Identity on Monocytes Colonizing the Liver Macrophage Niche

doi: 10.1016/j.immuni.2019.08.017

Figure Lengend Snippet:

Article Snippet: Recombinant Human/Mouse/Rat BMP-2 Protein , RnD Systems , Cat#355-BM-010.

Techniques: Control, Recombinant, Blocking Assay, Irradiation, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Microarray, Software, Microscopy

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 1. BMP15 and GDF9 serum ELISA development and validation. (a–d) Standards in the (a and c) BMP15 and (b and d) GDF9 ELISAs were recombinant pro-BMP15 and recombinant mature human GDF9 (GDF9), respectively. (a and b) Specificity was assessed using increasing doses of recombinant human proteins of the TGF-b superfamily (GDF5, GDF6, and GDF7). (c and d) Parallelism between the standards and serum samples was assessed in dose-response curves with pooled serum samples from women of reproductive age (serum QC-1 to 3, top dose 100 mL of serum) and (d) for GDF9, with pro-GDF9. Data are mean 6 SD of (a, b, and d) triplicates and (c) duplicates. OD, optical density.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Recombinant

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 2. BMP15 and GDF9 antibody validation. (a–d) Effect of neutralizing the detection and coating antibodies with specific (28A, 53-1, and 72B) and nonspecific (OVA) peptides on the pro-BMP15 and GDF9 standards and/or serum BMP15 QC (QC-4) in the (a and b) BMP15 and (c and d) GDF9 ELISAs. Absorbance was normalized to the standard/sample in the absence of neutralizing peptide and presented as a proportion (%) of the signal. Data are mean 6 SD of (a and c) triplicates and (b and d) duplicates.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Biomarker Discovery

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 3. Serum BMP15 and GDF9 in women at early, middle, and late reproductive age. (a, c, and e) BMP15 and (b, d, and f) GDF9 in 189 women, including asymptomatic women of reproductive age (n 5 6, 22 to 41 y of age), women being treated for infertility (n 5 154, 25 to 52 y of age), perimenopausal women (n 5 3, 48 to 52 y of age), and postmenopausal women (n 5 25, 48 to 66 y of age). Data are presented (a and b) as scatterplots with Spearman correlation coefficients, (c and d) stratified into 5-y age intervals (or 10-y intervals where n , 10 for that interval), or (e and f) as the proportion of women with serum BMP15 or GDF9 above the limit of quantification. (a–d) Dots represent individual women. (a and b) C, premenopausal women; s, postmenopausal women. (c and d) Black dots with bars represent the estimated geometric mean 6 95% CI, dashed horizontal lines indicate assay limit of quantification, and numbers above the x-axis show number of women. **P , 0.01.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 4. Follicular phase concentrations of serum BMP15 and GDF9 in women undergoing IVF. (a and c) Individual serum samples throughout GnRH antagonist FSH-stimulated cycles (n 5 15, plus an additional two women for BMP15). Black bars represent blood samples taken prior to exogenous FSH stimulation, and subsequent gray bars are samples taken every 2 to 3 d throughout FSH stimulation. Dashed horizontal lines indicate the assay limit of quantification, and samples with BMP15/GDF9 below this value are denoted “Undetectable.” (b) For women who had a blood sample prior to FSH stimulation (n 5 11), BMP15 concentrations of subsequent samples were normalized to the BMP15 of this baseline sample (day 2 to 3), shown as mean 6 SEM.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 5. Serum concentrations of BMP15, GDF9, and AMH and age of women relative to ovarian stimulation type. (a–f) Stimulated cycles were compared: GnRH antagonist and agonist (LDR and short Flare) ovarian stimulation, relative to (a and c) BMP15, (b and d) GDF9, (e) AMH, and (f) age. Data are represented (a, b, e, and f) as estimated geometric mean 6 95% CI (black dots with bars) with gray dots representing individual women, or (c and d) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. (a, b, e, and f) Dashed horizontal lines indicate the assay limit of quantification. Numbers above the x-axes show the number of women. *P , 0.05; ***P , 0.001.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 6. Serum concentrations of BMP15, GDF9, and AMH and age of stimulated and unstimulated women during infertility treatment. All stimulated cycles were combined and compared with unstimulated cycles, including natural cycle monitoring or IUI without FSH, hormone replacement therapy, or ovulation induction. Data are represented (a, b, e, and f) as estimated geometric mean 6 95% CI (black dots with bars), with gray dots representing individual women, or (c and d) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. Dashed horizontal lines indicate the assay limit of quantification (a, b, e, and f). Numbers above the x-axes show the number of women.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 7. Serum BMP15, GDF9 and AMH relative to the number of oocytes retrieved following ovarian stimulation. (a–f) Women with cancelled cycles due to too few follicles developed with stimulation and with increasing numbers of oocytes retrieved (binned 1 to 5, 6 to 10, 11 to 15, .15 oocytes) were compared with respect to BMP15, GDF9, and AMH. Data are represented as (a–c and g–i) estimated geometric mean 6 95% CI (black dots with bars), with gray dots representing individual women, or (d–f) as the proportion of women with serum BMP15 or GDF9 above the assay limit of quantification. Dashed horizontal lines indicate the assay limit of quantification (a–c and g–i). Numbers above the x-axes show number of women. ***P , 0.001.

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques:

Journal: Endocrinology

Article Title: Serum Concentrations of Oocyte-Secreted Factors BMP15 and GDF9 During IVF and in Women With Reproductive Pathologies.

doi: 10.1210/en.2019-00264

Figure Lengend Snippet: Figure 8. Relationship between serum biomarker concentration and oocyte number in women with and without polycystic ovaries and PCOS. (a–c) Regression modeling of (a) BMP15, (b) GDF9, and (c) AMH concentrations relative to the number of oocytes retrieved following ovarian stimulation in women without PCOS (n; solid regression lines) and women with PCOS (4; dashed regression lines). Tables below each figure indicate the relative change (Rel. change) in biomarker concentration associated with increasing number of oocytes in each patient group and associated P values. The P(int) term describes the significance of the interaction between

Article Snippet: Recombinant proteins used as standard preparations in the BMP15andGDF9ELISAswere promature human (h) BMP15 (proBMP15; which is a noncovalent complex–formed postproteolytic cleavage of the pro-domain and mature-domain of BMP15) and mature hGDF9 (R&D Systems, Minneapolis, MN), respectively.

Techniques: Biomarker Discovery, Concentration Assay