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: 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: Respiratory Research

Article Title: CircGSAP alleviates pulmonary microvascular endothelial cells dysfunction in pulmonary hypertension via regulating miR-27a-3p/BMPR2 axis

doi: 10.1186/s12931-022-02248-7

Figure Lengend Snippet: BMPR2 is the target of miR-27a-3p. A Volcano map analysis of RNA sequencing in PAECs overexpressing circGSAP. B miRWalk, StarBase, and TargetScan bioinformatics software predicted mRNAs that could bind to miR-27a-3p and the intersection with RNA sequencing results. C qRT–PCR was used to detect mRNA that could bind to miR-27a-3p in PMECs treated with circGSAP (n = 3). D Expression levels of BMPR2 in PMECs treated with circGSAP siRNA (n = 3). E , F Expression levels of BMPR2 in PMECs treated with miR-27a-3p mimics and inhibitor (n = 3). G Expression levels of BMPR2 in lung tissues of IPAH patients (n = 4). H Expression levels of BMPR2 in lung tissues of control, MCT, MCT-AAV-NC and MCT-AAV group rats (n = 11). I , J The protein levels of BMPR2 in circGSAP-treated and siRNA circGSAP-treated PMECs (n = 4). K Dual-luciferase assays were used to validate the interactions between miR-27a-3p and BMPR2 (n = 3). All data are presented as the mean ± SEM. Scale bar: 100 μm

Article Snippet: BMPR2 levels of PMECs under circGSAP regulation were determined using Human BMPR2 (Bone morphogenetic protein receptor type-2) ELISA Kit (EH4805, Wuhan Fine Biotech Co.) following the manufacturer’s protocol.

Techniques: RNA Sequencing Assay, Software, Quantitative RT-PCR, Expressing, Luciferase

Journal: Respiratory Research

Article Title: CircGSAP alleviates pulmonary microvascular endothelial cells dysfunction in pulmonary hypertension via regulating miR-27a-3p/BMPR2 axis

doi: 10.1186/s12931-022-02248-7

Figure Lengend Snippet: Effects of BMPR2 on the proliferation, migration and mortality of PMECs. A Expression levels of BMPR2 in hypoxia-treated PMECs (n = 4). B Expression levels of BMPR2 in PMECs treated with siRNA BMPR2 (n = 3). C–F Cell proliferation analysis, wound healing analysis, cell migration analysis and cell mortality analysis of PMECs transfected with siRNA BMPR2 under normoxia (n = 5 or 4). G–J Cell proliferation analysis, wound healing analysis, cell migration analysis and cell mortality analysis of PMECs cotransfected with miR-27a-3p inhibitor and siRNA BMPR2 under normoxia (n = 5 or 4). All data are presented as the mean ± SEM. Scale bar: 100 μm

Article Snippet: BMPR2 levels of PMECs under circGSAP regulation were determined using Human BMPR2 (Bone morphogenetic protein receptor type-2) ELISA Kit (EH4805, Wuhan Fine Biotech Co.) following the manufacturer’s protocol.

Techniques: Migration, Expressing, Transfection

Journal: Respiratory Research

Article Title: CircGSAP alleviates pulmonary microvascular endothelial cells dysfunction in pulmonary hypertension via regulating miR-27a-3p/BMPR2 axis

doi: 10.1186/s12931-022-02248-7

Figure Lengend Snippet: A schematic diagram illustrating the hypothetical model by which circGSAP adsorbed miR-27a-3p via a sponging mechanism and increased the BMPR2 signaling pathway, improving the overproliferation and migration, increasing mortality of PMECs in the remodeled pulmonary arteries

Article Snippet: BMPR2 levels of PMECs under circGSAP regulation were determined using Human BMPR2 (Bone morphogenetic protein receptor type-2) ELISA Kit (EH4805, Wuhan Fine Biotech Co.) following the manufacturer’s protocol.

Techniques: Migration