human bmpr2 elisa kit  (Elabscience Biotechnology)

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a ) Schematic diagram of sample processing for RNA sequencing. ( b ) Under the condition of BMRP2 knockdown, the stimulation of progesterone promoted the enrichment of pro-proliferative genes in PASMCs. ( c ) Both infecting shBMPR2 lentivirus (MOI=30, 48 hr) and transfecting siBMPR2 (50 nM, 48 hr) effectively decreased the protein level of BMPR2 in PASMCs. ( d ) Progesterone had little effect on the proliferation of normal PASMCs. ( e-h ) Under the condition of BMPR2 knockdown, progesterone (for 24 hr) significantly promoted the proliferation of PASMCs. (e) used Alarmar blue assay, (f-g) were EdU assay and its statistical graph, and (h) was the expression of PCNA. Abbreviation : PASMCs, pulmonary artery smooth muscle cells; NC, normal PASMCs; NC_C, normal PASMCs stimulated with 17β-estradiol; NC_Y, normal PASMCs stimulated with progesterone; KD, shBMRP2 lentivirus-infected PASMCs; P100, 100nM of progesterone; P1, 1μM of progesterone; PCNA, proliferating cell nuclear antigen. ns, non-significance; *, P <0.05; **, P <0.01; ****, P <0.001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: RNA Sequencing, Knockdown, EdU Assay, Expressing, Infection

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a ) Progesterone (24 hr) enriched the genes of “MAPK pathways” and “Pathways in cancer” in BMRP2-knockdown PASMCs. ( b ) Progesterone (for 2 hr) activated the ERK pathway in BMPR2-knockdown PASMCs. ( c ) Venn graph, which brought into all the differentially-expressed genes with the fold change > 1.4, showed that MYC and EDN1 were upregulated by progesterone (24 hr) only under the condition of BMPR2 knockdown. ( d-e ) Progesterone upregulated the mRNA and protein of c-MYC and EDN1 in BMPR2-knockdown PASMCs. Abbreviation : ERK, extracellular regulated protein kinases; EDN1, endothelin1; MAPK, mitogen-activated protein kinase. **, P <0.01; ****, P <0.001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Knockdown

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a ) Alarmar blue assay showed that PD0325901 and Bosentan (for 24 hr) reversed the pro-proliferative effects of progesterone in BMPR2-knockdown PASMCs . (b-c ) EdU assay showed that PD0325901 and Bosentan (for 24 hr) also reversed the effects of progesterone. ( d ) Inhibiting ERK pathway by PD0325901 reversed the upregulation of these proteins induced by progesterone in BMPR2-knockdown PASMCs. Abbreviation : ECE1, endothelin converting enzyme 1; PD, PD0325901; Bos, Bosentan. ns, non-significance; *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Knockdown, EdU Assay

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a-b ) Transfecting siPGR (200nM, 72 hr) effectively downregulated the mRNA and protein of PGR. ( c-e ) Knockdown of PGR reversed the pro-proliferative effects of progesterone (for 24 hr) in BMPR2-knockdown PASMCs. (c) used Alarmar blue assay, and (d-e) were EdU assay and its statistical graph. ( f ) Knockdown of PGR reversed the upregulation of these proteins induced by progesterone (for 2 hr) in BMPR2-knockdown PASMCs. Abbreviation: PGR, progesterone receptor. *, P <0.05; **, P <0.01; ***, P <0.001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Knockdown, EdU Assay

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a-b ) Progesterone (for 2 hr) promoted the nuclear translocation of c-JUN in BMPR2– knockdown PASMCs, which could be reversed by knockdown of PGR. ( c ) Nucleus and cytoplasm ratio of mean fluorescence intensity of c-JUN. ( d ) Compared with RLP30 (the positive control) and IgG (the negative control), the %input of cJUN was over 4%, demonstrating its binding to the promoter region of EDN1. ( e ) JASPAR database predicted that AP-1 (the dimer of c-JUN and c-FOS) could combine onto the promoter region of EDN1. Abbreviation : ChIP, chromatin immunoprecipitation; AP-1, activator protein-1. **, P <0.01; ***, P <0.001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Translocation Assay, Knockdown, Fluorescence, Positive Control, Negative Control, Binding Assay, Chromatin Immunoprecipitation

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a ) Alarmar blue assay showed that progesterone (24 hr) upregulated the proliferation in iPSCs-VSMCs of a HPAH patient, but not influenced iPSCs-VSMCs of a normal person. ( b-c ) Transwell assay showed that progesterone upregulated the migration (8 hr) in iPSCs-VSMCs of a HPAH patient, but not influenced iPSCs-VSMCs of a normal person. ( d ) Progesterone (2 hr) induced the phosphorylation of ERK in iPSCs-VSMCs. ( e ) In comparison with normal iPSCs-VSMCs, EDN1 was significantly increased in PAH iPSCs-VSMCs that could be further upregulated by progesterone (24 hr). Abbreviation: iPSCs, induced pluripotent stem cells; VSMCs, vascular smooth muscle cells; HPAH, hereditary pulmonary artery hypertension; BMPR2 mut, BMRP2-mutation carrier; WT, wild type. ns, non-significance; *, P <0.05.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Transwell Assay, Migration, Phospho-proteomics, Comparison, Mutagenesis

Journal: bioRxiv

Article Title: Progesterone is an Inducement of Heritable Pulmonary Arterial Hypertension with BMPR2 Mutation

doi: 10.1101/2023.04.21.537897

Figure Lengend Snippet: ( a ) Time flowchart of animal intervention and phenotype detection. ( b-c ) Right heart catheterization experiments showed the increase of RVSP in CKO and CKO+P group. (n=4 per group) ( d-e ) H&E stain showed the vascular remodeling of pulmonary arterioles (<50μM and 50-100μM) in CKO and CKO+P group and the statistical analyses (4 mice per group, 3 fields per mouse). ( f ) IF stain of vWF (green) and αSMA (red) showed the proliferation and hypertrophy of PASMCs in CKO and CKO+P group. Abbreviation: ♀, female; RVSP, right ventricle systolic pressure; CKO, SM22-cre BMPR2 flox +/- mice; P, progesterone; IF, immunofluorescence; vWF, von Willebrand factor; αSMA, smooth muscle actin. *, P <0.05; **, P <0.01; ****, P <0.0001.

Article Snippet: Human BMPR2 ELISA Kit (orb406355, Biorbyt, United Kingdom) was used to detect the level of soluble BMPR2 in serum.

Techniques: Staining, Immunofluorescence

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

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