bmpr1b antibody  (Novus Biologicals)

Journal: bioRxiv

Article Title: Tunneling nanotubes propagate a BMP-dependent preneoplastic state

doi: 10.1101/2025.11.11.687852

Figure Lengend Snippet: | Actors of the BMP signaling pathway are transferred to acceptor cells through TNTs. a, GOBP subset from a list of 6 protein hits dysregulated between NoTf accept and NoTf no accept cells, generated through STRING from proteins of the BMP and p38 pathways, chosen from the literature. The size of each circle represents the number of genes in the respective category; the intensity of the color represents the P-value, determined by a hypergeometric test. b, Box and whiskers plots of Log 2 LFQ values (Protein Levels) of ID1 ( ID1 ), Nup214 ( NUP214 ), Spartin ( SPART ), ATF2 ( ATF2 ), Rac1 ( RAC1 ) and desmoglein-3 ( DSG3 ), for indicated sorted cell populations. Statistical significance was calculated using one-way ANOVA with Dunnett’s multiple comparison test (n = 5 biological replicates; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). For clarity, only comparison between NoTf and NoTf accept cells is shown. c, Immunofluorescence images of actin (red) and BMPR1b (green) in indicated cells. Nuclei (blue). Scale bars, 20 μm. d, Correlative light scanning electron microscopy (SEM) of BMPR1b, immunostained with Alexa Fluor ® 488-Fluoronanogold ™ (green) in M1B26 cells. From left to right: confocal, SEM, zooms, correlative (overlay). Scale bars, 10 μm, unless otherwise indicated. e, Immunofluorescence of actin (green), after transfection with siRNA pools targeting BMPR1b (siBMPR1b) or GPER (siGPER). Non-targeting siRNA: siCTRL. f,g, BMPR1B and GPER transcript (f) and protein expression upon their silencing (f),with siCTRL set at 1 (n=4), and TNT number (> 8 μm) per cell and % of long TNTs (> 50 μm) upon indicated silencing, normalized against siCTRL (g). Error bars depict SD. Statistical significance was calculated using a Student’s two-sided t test with Welch’s correction (n = 4 biological replicates, *** p < 0.001). h, Schematic of experimental design: after sorting, cell populations were either lysed and subjected to RT-qPCR for selected transcript analyses, or grown for several weeks with or without BMP2/IL6, and immunoblot analyses performed at end points. i, RT-qPCR measurement of BMPR1b mRNA in indicated cell populations, the expression in NoTf cells set at 1. Statistical significance was calculated using one-way ANOVA with post-hoc Tukey test (n = 5 biological replicates). For clarity, only comparison between NoTf accept and NoTf cells is shown (**** p < 0.0001).

Article Snippet: After PBS washing, primary antibody against BMPR1b (1:300 #sc-293428 Santa Cruz) and its corresponding isotype were incubated overnight at 4°C.

Techniques: Generated, Comparison, Immunofluorescence, Electron Microscopy, Transfection, Expressing, Quantitative RT-PCR, Western Blot

Journal: bioRxiv

Article Title: Tunneling nanotubes propagate a BMP-dependent preneoplastic state

doi: 10.1101/2025.11.11.687852

Figure Lengend Snippet: | Non-transformed cells acceptor of information via TNTs from transformed cells display preneoplastic features. a, Left, quantification of BMPR1b immunoblots performed on NoTf accept cells grown in the presence of BMP2/IL6, for indicated weeks post sorting. Box and whiskers plot of 6 biological replicates. Statistical significance was calculated using a one-way ANOVA with non-parametric Kruskal-Wallis test, and for clarity, only p-values ≤ 0.05 are indicated. Right, quantification of BMPR1b immunoblots performed on indicated cells, 14 weeks post sorting, with NoTf conditions set at 1 (n = 6 biological replicates). Statistical significance was calculated using a one-way ANOVA with post-hoc Tukey test (*** p < 0.001). b,c, Quantification of p-SMAD1/5/8/SMAD1/5/8 (b) and p-p38/p38 (c) immunoblots performed on NoTf accept cells grown in the presence of BMP2/IL6, for indicated weeks post sorting. Box and whiskers plot of 6 biological replicates. Statistical significance was calculated using a one-way ANOVA with non-parametric Kruskal-Wallis test, and for clarity, only p-values ≤ 0.05 are indicated. d, RNA-sequencing results of transcriptomic expression levels of HOXB6, CCND2, EREG and HRAS in indicated cell lines (n = 3 biological replicates). Data deposited on the Gene Expression Omnibus repository, GSE186734 . Error bars depict SD. Statistical significance was calculated using a one-way ANOVA with post-hoc Tukey test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). e, RT-qPCR measurements of HOXB6, CCND2, EREG and HRAS mRNA in indicated cell populations, with the expression in NoTf cells set at to 1. Error bars depict SD. Statistical significance was calculated using a one-way ANOVA with post-hoc Tukey test (Biological replicates: n = 8 for HOXB6, n = 5 for CCND2, n = 7 for EREG, n = 8 for HRAS). For clarity, only comparison between NoTf accept and NoTf cells is shown (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). f, Schematic of long-term soft agar colony formation assays. g, Quantification of soft agar clones of indicated sorted cells, after 16 weeks in culture with or without BMP2/IL6 (n = 3 biological replicates). Error bars represent SD. Statistical significance was calculated using a one-way Welch and Brown-Forsythe ANOVA test (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). For clarity, only p-values ≤ 0.05 are indicated, and comparisons with Tf cells are omitted. h, Model depicting how TNTs are involved in the propagation of a BMP-dependent preneoplastic state. Fibrils and fibers depict extracellular matrix components.

Article Snippet: After PBS washing, primary antibody against BMPR1b (1:300 #sc-293428 Santa Cruz) and its corresponding isotype were incubated overnight at 4°C.

Techniques: Transformation Assay, Western Blot, RNA Sequencing, Expressing, Gene Expression, Quantitative RT-PCR, Comparison, Clone Assay

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: Genotyping was performed by Taqman method using DNA extracted from peripheral blood leukocytes. A. Representative fluorescent signal plots from genotyping of SNPs in BMPR1B . B. Through linkage disequilibrium evaluation, D ' and r 2 values were plotted at top right and bottom left triangles, respectively, for normal controls (upper panel) and endometriosis patients (lower panel). Scales beneath the charts show the relative location of each SNP on chromosome 4. SNP1: rs1970801; SNP2: rs1434536; SNP3: rs11097457.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: Genotype and allele distributions of SNPs surrounding miRNA target site of BMPR1B in endometriosis patients and controls.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: Association between haplotypes of rs1970801 and rs1434536 in BMPR1B with endometriosis and CA125 level in patients.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: A. Serum CA125 levels in endometriosis patients were grouped by their rs1434536 genotypes. Bold lines indicate median values of CA125 levels in each genotype group. Notch regions define 95% confidence interval estimations of median values. B. Gene expression levels of BMPR1B and CA125 (MUC16) in ectopic endometrial tissues were measured by quantitative PCR for correlation study. C. Immunohistochemistry (IHC) staining was performed on ectopic endometrial tissues from patients. Anti-CD10 staining, a stroma cell-specific marker, serves as the reference to normalize immunointensity. Ectopic endometrium samples represented patients with rs1434536 genotype of CC, CT; and TT, as labeled on top of each column. A total of 6, 8 and 12 samples from tissue microarrays could be defined as TT, CT and CC genotype groups, respectively, and their expression levels for BMPR1B and CA125 were scored as described in method section (right panel). Box plot and ANOVA P value calculation were performed in R software suite.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunohistochemistry, Staining, Marker, Labeling, Software

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: A. Luciferase reporter assay was employed to evaluate repression of BMPRIB in endometrial cell lines with C (HEC-1-A) or T (RL95-2) allele at rs1434536. Luciferase reporter pGL4.73 vectors, containing the reference C or variant T bearing sequence of BMPR1B 3′UTR, were transiently introduced into both cell lines. At 48h post-transfection, luciferase activities were measured and normalized to phosphatase activities. B. Cell growth of HEC-1-A cells transfected with empty vector or vector containing miR-125b-wt or miR-125b-mt with a mutated base complementary to the minor allele at rs1434536. At different time intervals, cells were counted using hemocytometer. C. HEC-1-A cells transfected with vectors containing miR-125b-wt or miR-125b-mt were counted and rated with reference to total positively transfected (GFP+) cells. D. Expression levels of known BMPR1B responsive genes were analyzed in HEC-1 cells transfected with miR-125b-wt or miR-125-mt constructs, using empty vector-treated cells as references. E. Expression levels of IL-1β in clinical samples were measured by quantitative PCR. Statistical significance is calculated by Student t-test: *, P <0.05; **, P <0.01.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques: Luciferase, Reporter Assay, Variant Assay, Sequencing, Transfection, Plasmid Preparation, Expressing, Construct, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: BMPR1B Up-Regulation via a miRNA Binding Site Variation Defines Endometriosis Susceptibility and CA125 Levels

doi: 10.1371/journal.pone.0080630

Figure Lengend Snippet: Mature miRNAs including miR-125b are produced by several processing steps where the microprocessor complex (Drosha/DGCR8), exportin 5 and Dicer/TRBP/PACT complex have been known to involve in. The resulting miR-125b forms a RISC complex with Ago and silences BMPR1B gene by binding to the complementary sequence in BMPR1B 3′UTR. Impaired recognition due to genetic variations in the mir-125b seed region reduces suppressive effect of mir-125b, resulting in up-regulation of BMPR1B. The CA125 level, a biomarker for endometriosis and ovarian cancer, was found reversely correlated with BMPR1B in endometrium cells. Elevated BMPR1B levels in endometrium cells have been proven to reduce cell proliferation and migration activity via down-regulation of IL-1β, indicating a lower risk to develop endometriosis.

Article Snippet: After cooling, sections were incubated with PBS for 60 minutes, followed by anti-BMPR1B antibody (Sigma-Aldrich), anti-CA125 antibody (Sigma-Aldrich) and anti-CD10 antibody (Sigma-Aldrich) at 1∶50 dilution overnight at 4°C.

Techniques: Produced, Binding Assay, Sequencing, Biomarker Assay, Migration, Activity Assay

Journal: Journal of Zhejiang University. Science. B

Article Title: Cathepsin D knockdown regulates biological behaviors of granulosa cells and affects litter size traits in goats

doi: 10.1631/jzus.B2100366

Figure Lengend Snippet: Real-time PCR primer details

Article Snippet: The membranes were incubated with primary antibodies, including anti-CTSD (1:2000; Bioss, bs-1615R, Beijing, China), anti-BMPR-IB (1:1500; Bioss, bs-6639R), anti-FSHR (1:1500; Bioss, bs-0895R), anti-INHA (1:1000, Bioss, bs-1032R), and anti-‍β‍-‍actin (1:3000; Affinity Biosciences, AF7018, USA) ( Tang et al., 2019 ).

Techniques: Real-time Polymerase Chain Reaction, Sequencing, Amplification