Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 reduces BMPR2-ECD ligand-binding activity in a modified immunoprecipitation assay. Neutralizing the ligand-binding activity of BMPR2-ECD using various amounts of 3F6. Results are quantified by ELISA and expressed as mean ± SEM relative to the ligand-binding activity of BMPR2-ECD in the absence of 3F6. n ≥ 3 per condition. Asterisk indicates p < 0.05 by paired t test

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Ligand Binding Assay, Activity Assay, Modification, Immunoprecipitation, Enzyme-linked Immunosorbent Assay

Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 reduces activation of BMP pathway in HEK293T cells. a Expression of endogenous BMPR2 by HEK293T cells compared to β-actin loading control. Approximate molecular weights are indicated. b , c BMP2 induces phosphorylation of SMAD1, 5, and 8 (pSMAD1,5,8) in HEK293T cells and this response is blunted by pre-treatment with 3F6. Approximate molecular weights are indicated in b . Results are quantified in c and expressed as mean ± SEM ratio of phosphorylated SMAD1,5,8: total SMAD1 relative to BMP2 treatment alone. No effect on BMP2-responsiveness was observed with pre-treatment using control ascites (Ctrl Asc.). n = 3 per condition. Asterisk indicates p < 0.05 by paired t test

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Activation Assay, Expressing

Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 has no effect on BMP-responsiveness in BMPR2 knock-down HEK293T cells. a Expression levels of endogenous BMPR2 in scramble control HEK293T cells (Control) and HEK293T cells carrying anti- BMPR2 shRNA ( BMPR2 -KD) compared to HPRT1 loading control. Data are expressed as normalized to scramble control (Control) using the 2 −∆∆Ct method. n = 6 per condition. Asterisk indicates p < 0.05 by unpaired t test. b , c Expression of endogenous BMPR2 in scramble control HEK293T cells (Control) and BMPR2 -KD HEK293T cells compared to β-actin loading control. Approximate molecular weights are indicated in b . Representative immunoblot is shown in b and results from three independent runs are quantified in c (data are expressed as mean ± SEM ratio of BMPR2: β-actin normalized to scramble control (Relative Expression)). Asterisk indicates p < 0.05 by paired t test. d , e BMP2 induces phosphorylation of SMAD1, 5, and 8 (pSMAD1,5,8) in BMPR2 - KD HEK293T cells. No effect on BMP2-responsiveness of BMPR2 - KD HEK293T cells was observed with pre-treatment using control ascites (Ctrl Asc.) or 3F6. Approximate molecular weights are indicated in d . Results are quantified in E and expressed as mean ± SEM ratio of phosphorylated SMAD1,5,8: total SMAD1 relative to BMP2 treatment alone (Relative pS1:S1). n = 4 per condition

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Expressing, shRNA, Western Blot

Journal: Respiratory research

Article Title: Altered maturation and activation state of circulating monocytes is associated with their enhanced recruitment in pulmonary arterial hypertension.

doi: 10.1186/s12931-025-03182-0

Figure Lengend Snippet: Fig. 6 Functional studies related to loss of BMPR2 in human monocytes. Induced pluripotent stem cells (iPSC) derived from HPAH patients or Controls were differentiated into induced monocytes (iMono) using a stepwise differentiation protocol outlined in the Methods. iMono of HPAH patients and Controls were compared for (A) CD14 cell surface expression, assessed by FACS; (B) Number of iMono per mL. on day 19 of differentiation. (C)STAT1 gene expression by RT-qPCR. (D) Schematic demonstrating iEC and iMono co-culture approach for assessing adhesion. iMono adhesion to Control or HPAH iEC, per field of view. (E) Schematic demonstrating iEC and iMono co-culture approach for assessing transendothelial migration. Migrated of iMono across Control or HPAH iEC. (F) VE-cadherin and ICAM1 expression in co-culture of iMono and iEC with quantification based on mean fluorescent intensity (MFI) of 10 FOV for each replicate. Representative images of the iEC/iMono co-culture including phase, DAPI (blue) VE-cadherin (CDH5) (green) and ICAM1 (red). Red dotted outline on phase and DAPI images show adhered iMono. White dotted outline on merged images show corresponding iMono (pink). Bars represent mean ± SEM. For all experiments, n = 3 biological replications. + denotes the minimum achievable P-value for n = 3 was reached by the non-parametric t test

Article Snippet: SiRNA BMPR2 knockdown in THP1 cell line THP1 cells at a density of 5 × 105 cells/mL were cultured overnight in a 6-well plate, followed by siRNA transfection with siRNA BMPR2 (SMARTpool ON-TARGETplus, Origene, Rockville, MD, Cat#: L-001230-00-005) or siRNA Control (ON-TARGETplus non-targeting Control siRNA, Origene, Cat#: D-001230-01) using the TransIT-TKO reagent (Mirus, Madison, WI Cat# MIR2150) according to the manufacturer’s recommendations.

Techniques: Functional Assay, Derivative Assay, Expressing, Gene Expression, Quantitative RT-PCR, Co-Culture Assay, Control, Migration

Journal: Genes

Article Title: Reduction of BMPR2 mRNA Expression in Peripheral Blood of Pulmonary Arterial Hypertension Patients: A Marker for Disease Severity?

doi: 10.3390/genes13050759

Figure Lengend Snippet: Baseline characteristics of PAH patients.

Article Snippet: Serum from each proband was pipetted into a 96-well plate and treated with ELISA reagents according to the manufacturer’s instructions (human BMPR2 ELISA kit, Elabscience, Wuhan, China).

Techniques: Variant Assay, Biomarker Discovery, Filtration, Diffusion-based Assay

Journal: Genes

Article Title: Reduction of BMPR2 mRNA Expression in Peripheral Blood of Pulmonary Arterial Hypertension Patients: A Marker for Disease Severity?

doi: 10.3390/genes13050759

Figure Lengend Snippet: Recruitment of study population from May 2019 to January 2020. 110 participants were screened, and samples were collected of which 109 were analyzed and included. BMPR2 : bone morphogenetic protein receptor type II, PAH: pulmonary arterial hypertension, PVOD: pulmonary veno-occlusive disease.

Article Snippet: Serum from each proband was pipetted into a 96-well plate and treated with ELISA reagents according to the manufacturer’s instructions (human BMPR2 ELISA kit, Elabscience, Wuhan, China).

Techniques:

Journal: Genes

Article Title: Reduction of BMPR2 mRNA Expression in Peripheral Blood of Pulmonary Arterial Hypertension Patients: A Marker for Disease Severity?

doi: 10.3390/genes13050759

Figure Lengend Snippet: Mean relative BMPR2 mRNA expression in whole blood. The boxplots provide median values (horizontal lines), interquartile range (box), 1.5× interquartile range (whiskers), outliers (indicated by circles within 1.5 to 3× interquartile range) and extreme outliers (indicated by asterisks > 3× interquartile range). A significant difference of BMPR2 mRNA expression could be identified between healthy controls, BMPR2 non-carriers and BMPR2 variant carriers. With Bonferroni correction, p -values were healthy controls vs. non-carriers p = 0.453, non-carriers vs. variant carriers p = 0.0002, and healthy controls vs. variant carriers p < 0.0001. The 1/delta cycle threshold (1/∆CT) denotes the level of BMPR2 mRNA gene expression measured by qPCR. n.s. = non-significant.

Article Snippet: Serum from each proband was pipetted into a 96-well plate and treated with ELISA reagents according to the manufacturer’s instructions (human BMPR2 ELISA kit, Elabscience, Wuhan, China).

Techniques: Expressing, Variant Assay, Gene Expression

Journal: Genes

Article Title: Reduction of BMPR2 mRNA Expression in Peripheral Blood of Pulmonary Arterial Hypertension Patients: A Marker for Disease Severity?

doi: 10.3390/genes13050759

Figure Lengend Snippet: Correlation of BMPR2 mRNA expression with laboratory and clinical characteristics.

Article Snippet: Serum from each proband was pipetted into a 96-well plate and treated with ELISA reagents according to the manufacturer’s instructions (human BMPR2 ELISA kit, Elabscience, Wuhan, China).

Techniques: Expressing, Filtration

Journal: European Journal of Pharmacology

Article Title: Sex-dependent right ventricular hypertrophic gene changes after methamphetamine treatment in mice

doi: 10.1016/j.ejphar.2021.174066

Figure Lengend Snippet: TaqMan® gene expression assays IDs used for the gene expression experiments.

Article Snippet: Bmpr2 [bone morphogenetic protein receptor, type II (serine/threonine kinase)] , Mm00432134_m1.

Techniques: Gene Expression, Ubiquitin Proteomics

Journal: European Journal of Pharmacology

Article Title: Sex-dependent right ventricular hypertrophic gene changes after methamphetamine treatment in mice

doi: 10.1016/j.ejphar.2021.174066

Figure Lengend Snippet: Expression of genes associated with PAH and fibrosis in lung tissue isolated from A) vehicle- (white) and methamphetamine- (black) treated female mice and B) vehicle- (white) and methamphetamine- (black) treated male mice. BMPR2 (Bone Morphogenetic Protein Receptor Type II), HTR1B (5-Hydroxytryptamine (Serotonin) Receptor 1B), TGFβR1 (Transforming Growth Factor Beta Receptor I), ESR 1 (Estrogen Receptor Alpha), ESR 2 (Estrogen Receptor Beta), GPER (G Protein-Coupled Estrogen Receptor 1), TGFβ1 (Transforming Growth Factor Beta 1), CYP1A1 (Cytochrome P450 Family 1 Subfamily A polypeptide 1),CYP1B1 (Cytochrome P450 Family 1 Subfamily B, Polypeptide 1), Col1a1 (collagen type I), Col3a1 (collagen type III), FN1 (fibronectin 1). Results are expressed as mean ± standard error of the mean (S.E.M) (n = 6). *P < 0.05 vs. vehicle group.

Article Snippet: Bmpr2 [bone morphogenetic protein receptor, type II (serine/threonine kinase)] , Mm00432134_m1.

Techniques: Expressing, Isolation

Journal: European Journal of Pharmacology

Article Title: Sex-dependent right ventricular hypertrophic gene changes after methamphetamine treatment in mice

doi: 10.1016/j.ejphar.2021.174066

Figure Lengend Snippet: Expression of genes associated with PAH and fibrosis in the RV tissue isolated from A) vehicle- (white) and methamphetamine- (black) treated female mice and B) vehicle- (white) and methamphetamine- (black) treated male mice. ANP (natriuretic peptide type A), BNP (natriuretic peptide type B), BMPR2 (Bone Morphogenetic Protein Receptor Type II), HTR1B (5-Hydroxytryptamine (Serotonin) Receptor 1B), TGFβR1 (Transforming Growth Factor Beta Receptor I), TGFβ1 (Transforming Growth Factor Beta 1), Col1a1 (collagen type I), Col3a1 (collagen type III), FN1 (fibronectin 1), Smad 2 (SMAD family member 2), Smad 3 (SMAD family member 3), Smad 7 (SMAD family member 7), Smurf 1 (SMAD specific E3 ubiquitin protein ligase 1), Smurf 2 (SMAD specific E3 ubiquitin protein ligase 2). Results are expressed as mean ± standard error of the mean (S.E.M) (n = 6). *P < 0.05 vs. vehicle group.

Article Snippet: Bmpr2 [bone morphogenetic protein receptor, type II (serine/threonine kinase)] , Mm00432134_m1.

Techniques: Expressing, Isolation, Ubiquitin Proteomics

Journal: Gastroenterology

Article Title: BMPR2 as a Novel Predisposition Gene for Hereditary Colorectal Polyposis.

doi: 10.1053/j.gastro.2023.03.006

Figure Lengend Snippet: Figure 1. The BMP pathway in the colon. BMP2 and BMP4 are the most abundant ligands of the BMP pathway in the bowel mucosa. They are able to bind the BMPR1A-BMPR2 complex on the surface of colonocytes, therefore, triggering R-SMAD phosphorylation and downstream gene regulation in the canonical pathway. In addition, BMP can also mediate proliferation by convergent pathways, such as ERK, JNK, and p38. Created with BioRender.com.

Article Snippet: The identified BMPR2 variants were introduced into the BMPR2 coding sequence (no. 116718, Addgene) by a simple 1- step PCR amplification with nonoverlapping primers (Q5 SiteDirected Mutagenesis Kit, New England Biolabs, Ipswich, MA).

Techniques: Phospho-proteomics

Journal: Gastroenterology

Article Title: BMPR2 as a Novel Predisposition Gene for Hereditary Colorectal Polyposis.

doi: 10.1053/j.gastro.2023.03.006

Figure Lengend Snippet: Figure 2. BMPR2 variants drive a proliferative response and impair the BMP4-mediated growth inhibition. Characterization of the proliferative capacity of 2 different BMPR2 clones (B26 and B31) in which the 3 BMPR2 variants were reintroduced. (A) MTS cell proliferation assay. Samples were assayed in triplicate and the experiment was repeated 5 times (n ¼ 5). (B) Plating efficiency (colonies originated from single cells) in the colony-formation assay. Samples were assayed in duplicate and the experiment was repeated 3 times (n ¼ 3). (C) MTS cell proliferation assay in the presence or absence of 50 ng/mL of BMP4. Data are represented as the growth difference between treated and untreated cells. Samples were assayed in triplicate and the experiment was repeated 4 times (n ¼ 4). Data represent mean ± SD. *P < .05, **P < .01, ***P < .001 for the analysis of variance with the least significant difference post-hoc test.

Article Snippet: The identified BMPR2 variants were introduced into the BMPR2 coding sequence (no. 116718, Addgene) by a simple 1- step PCR amplification with nonoverlapping primers (Q5 SiteDirected Mutagenesis Kit, New England Biolabs, Ipswich, MA).

Techniques: Inhibition, Clone Assay, Proliferation Assay, Colony Assay

Journal: Gastroenterology

Article Title: BMPR2 as a Novel Predisposition Gene for Hereditary Colorectal Polyposis.

doi: 10.1053/j.gastro.2023.03.006

Figure Lengend Snippet: Figure 3. BMPR2 variants act through SMAD or non-SMAD effectors, depending on their position. Characterization of the SMAD1/5/8 pathway activity of 2 different BMPR2 clones (B26 and B31) in which the 3 BMPR2 variants were reintroduced. (A) Quantitative analysis of the phosphorylated levels of SMAD1 (Ser463/465) by means of enzyme-linked immunosorbent assay in the presence or absence of 50 ng/mL of BMP4. Samples were assayed in duplicate and the experiment was repeated 3 times (n ¼ 3). (B) Real-time PCR quantification of ID1 and ID3 expression levels, 2 canonical BMP-SMAD downstream targets, in clone B26. Samples were assayed in triplicate and the experiment was repeated 3 times (n ¼ 3). Data represent mean ± SD. *P < .05, **P < .01, ***P < .001 for the analysis of variance with the least significant difference post-hoc test.

Article Snippet: The identified BMPR2 variants were introduced into the BMPR2 coding sequence (no. 116718, Addgene) by a simple 1- step PCR amplification with nonoverlapping primers (Q5 SiteDirected Mutagenesis Kit, New England Biolabs, Ipswich, MA).

Techniques: Activity Assay, Clone Assay, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing