Journal: Chinese Medicine

Article Title: BMP6, a potential biomarker of inflammatory fibrosis and promising protective factor for dilated cardiomyopathy

doi: 10.1186/s13020-025-01062-9

Figure Lengend Snippet: Significant gene and assessment of biomarker sensitivity. A Heatmap of differential expression of immune-related genes between DCM and NC. B Heatmap of differential expression of senescene-related genes between DCM and NC. C Intersection of ImmDEGS, SenDEGS and gene sets in MEyellow. D Sensitivity and specificity analysis of ROC curves between BMP6 and DCM. E Correlation between BMP6 and 22 types of immune cells

Article Snippet: Additionally, the expression levels of proteins, including BMP6 (M06924-1, Boster, USA), SMAD3 (25494-1-AP, ProteinTech, USA), SMAD6 (sc-25321, Santa Cruz, USA), IL-1β (16806-1-AP, ProteinTech, USA), IL-18 (60070-1-Ig, ProteinTech, USA), IL-6 (21865-1-AP, ProteinTech, USA), TNF-α (17590-1-AP, ProteinTech, USA), and COL1A1 (sc-293182, Santa Cruz, USA), were detected using western blotting.

Techniques: Biomarker Discovery, Quantitative Proteomics

Journal: Chinese Medicine

Article Title: BMP6, a potential biomarker of inflammatory fibrosis and promising protective factor for dilated cardiomyopathy

doi: 10.1186/s13020-025-01062-9

Figure Lengend Snippet: BMP6 expression in different cellular subpopulations. A Differential expression of BMP6 in various cell types of the heart. B Differential gene expression in fibroblasts between DCM and NH. C Differential gene expression in endothelial cells between DCM and NH. D Elbow diagram obtained using the harmony method and we selected 0–15 as the number of dimensions. E Specific markers and expression levels in fibroblasts and myofibroblasts. F – I subdividing and annotating fibroblast subpopulations by TSNE and Umap methods. J BMP6 expression between conventional fibroblasts and myofibroblasts. K differential expression of BMP6 in fibroblasts between DCM and NH. L Specific markers and expression levels of macrophage subpopulations. M – P Subdividing and annotating macrophage subpopulations by TSNE and Umap methods. Q Differences in the percentage of macrophage subpopulations abundance between DCM and NH

Article Snippet: Additionally, the expression levels of proteins, including BMP6 (M06924-1, Boster, USA), SMAD3 (25494-1-AP, ProteinTech, USA), SMAD6 (sc-25321, Santa Cruz, USA), IL-1β (16806-1-AP, ProteinTech, USA), IL-18 (60070-1-Ig, ProteinTech, USA), IL-6 (21865-1-AP, ProteinTech, USA), TNF-α (17590-1-AP, ProteinTech, USA), and COL1A1 (sc-293182, Santa Cruz, USA), were detected using western blotting.

Techniques: Expressing, Quantitative Proteomics, Gene Expression

Journal: Chinese Medicine

Article Title: BMP6, a potential biomarker of inflammatory fibrosis and promising protective factor for dilated cardiomyopathy

doi: 10.1186/s13020-025-01062-9

Figure Lengend Snippet: Mendelian randomization analysis between BMP6 and DCM. A Scatter trend plot of effective SNPs. B The black line represents the nature of each SNP, and the red lines represent the combined nature of the SNPs analyzed by the IVW and MR Egger methods, respectively. C SNPs distribution map. D Leave-one-out analysis. The red line shows the results of the combined analysis

Article Snippet: Additionally, the expression levels of proteins, including BMP6 (M06924-1, Boster, USA), SMAD3 (25494-1-AP, ProteinTech, USA), SMAD6 (sc-25321, Santa Cruz, USA), IL-1β (16806-1-AP, ProteinTech, USA), IL-18 (60070-1-Ig, ProteinTech, USA), IL-6 (21865-1-AP, ProteinTech, USA), TNF-α (17590-1-AP, ProteinTech, USA), and COL1A1 (sc-293182, Santa Cruz, USA), were detected using western blotting.

Techniques:

Journal: Chinese Medicine

Article Title: BMP6, a potential biomarker of inflammatory fibrosis and promising protective factor for dilated cardiomyopathy

doi: 10.1186/s13020-025-01062-9

Figure Lengend Snippet: Immunofluorescence staining and cardiac tissue protein assay and BMP6 knockdown. A Immunofluorescence staining of markers specific to macrophages and type 2 macrophages. B and C Western blot was used to detect the changes in protein levels in the cardiac tissues between DCM and NC (n = 3). D and E The knockdown of BMP6 and the expression levels of SMAD6 and COL1A1 (n = 3). The siRNA group represents the knockdown group, si-NC represents the empty vector group, and blank represents the control group. The significance of * P in the above graph: * P < 0.05; ** P < 0.01. F Potential interventional herbs based on BMP6 inverse prediction

Article Snippet: Additionally, the expression levels of proteins, including BMP6 (M06924-1, Boster, USA), SMAD3 (25494-1-AP, ProteinTech, USA), SMAD6 (sc-25321, Santa Cruz, USA), IL-1β (16806-1-AP, ProteinTech, USA), IL-18 (60070-1-Ig, ProteinTech, USA), IL-6 (21865-1-AP, ProteinTech, USA), TNF-α (17590-1-AP, ProteinTech, USA), and COL1A1 (sc-293182, Santa Cruz, USA), were detected using western blotting.

Techniques: Immunofluorescence, Staining, Knockdown, Western Blot, Expressing, Plasmid Preparation, Control

Journal: Cell Death & Disease

Article Title: RNF4~RGMb~BMP6 axis required for osteogenic differentiation and cancer cell survival

doi: 10.1038/s41419-022-05262-1

Figure Lengend Snippet: A Media transfer restores differentiation of RNF4-deficient stalled progenitors. OD assisted by transfer of culture media from the indicated cells for 21 days of the indicated hBMSCs, as analyzed by AR staining. MT: transfer of culture media collected from wild-type hBMSCs (MT−) or hBMSCs undergoing OD (MT+) (see Methods for details). sh-Scr denotes scrambled control shRNA; sh-RNF4 denotes sh-RNF4. B Alkaline phosphatase (ALP) activity of the indicated hBMSCs under experimental conditions similar to A . C , D Identification of RNF4-regulated genes during OD and in hBMCs. C Schematic diagram of RNA-seq results of OD in control hBMSCs, or RNF4-deficient hBMSCs; OB osteoblasts. D Schematic diagram of RNA-seq results of control hBMSCs or RNF4-deficient hBMSCs, lower panels depict key GO groups of RNF4-regulated genes in hBMSCs. E , F qPCR analysis of BMP6 ( E ) and RGMb ( F ) mRNA levels in the indicated hBMSCs. BMP6 levels were determined 2 weeks after OI and RGMb levels in undifferentiated progenitors. sh-SCR; scrambled shRNA, control) n = 3 **** p < 0.0001; *** p < 0.001; * p < 0.1. Statistics was calculated by one-step ANOVA Graph-prism 6.

Article Snippet: The antibodies used were: anti-RNF4 (Merck, HPA022047) and BMP6 (Merck, HPA062683).

Techniques: Staining, shRNA, Activity Assay, RNA Sequencing Assay

Journal: Cell Death & Disease

Article Title: RNF4~RGMb~BMP6 axis required for osteogenic differentiation and cancer cell survival

doi: 10.1038/s41419-022-05262-1

Figure Lengend Snippet: A , B OD for 21 days of the indicated hBMSCs deficient in either BMP6 [shBMP6 ( A ) or RGMb (shRGMb) ( B ). OD analyzed by Alizarin Red (AR) staining. Cell viability was visualized using Brilliant violet (BV) and osteogenic induction is marked by “OI+”. C , D OD of hBMSCs in which RNF4 was knocked down using the indicated shRNF4. Where indicated, the differentiation media was supplemented with recombinant purified BMP6 or RGMb, or both BMP6 and RGMb proteins as indicated. E Alkaline phosphatase activity (Alk-Phos) in the indicated cells 21 days after OI. n = 3; ** P < 0. 01 * P < 0.1 Statistics was calculated by one-step ANOVA Graph-prism 6.

Article Snippet: The antibodies used were: anti-RNF4 (Merck, HPA022047) and BMP6 (Merck, HPA062683).

Techniques: Staining, Recombinant, Purification, Activity Assay

Journal: Cell Death & Disease

Article Title: RNF4~RGMb~BMP6 axis required for osteogenic differentiation and cancer cell survival

doi: 10.1038/s41419-022-05262-1

Figure Lengend Snippet: A Box-plot of RNF4 mRNA expression derived from publicly available human sarcoma samples. Sample size, “ n ”, is indicated in brackets. For the mixed osteosarcoma expression set by Aqelian, as well as the Mixed Ewing Sarcoma dataset by Savola (highlighted in red boxes), RNF4 expression in normal tissue and sarcoma samples are presented individually in ( B ). Data generated with the online R2 visualization tool, Genomics Analysis, and Visualization Platform ( https://hgserver1.amc.nl/cgi-bin/r2/main.cgi ). B Individual RNF4 mRNA expression levels of samples within the mixed osteosarcoma expression set by Aqelian (Left panel; red = normal, green = tumor) and the Mixed Ewing Sarcoma dataset by Savola (right panel; diagnosis: blue = normal, green = Ewing, red = askin, yellow = pnet, purple = non-classified; state: green = primary, red = metastasis, blue = recurrent; status: green = diseased, red = no evidence of disease (ned), red = alive with disease (awd)). Data were generated with the online R2 visualization tool, Genomics Analysis and Visualization Platform ( https://hgserver1.amc.nl/cgi-bin/r2/main.cgi ) and adapted to present normal vs. tumor samples. The green dotted line separates the normal and tumor samples. C , D Representative immuno-staining using antibodies against RNF4 and BMP6 on human pan-sarcoma TMA ( n = 30). In each set, normal non-tumor tissue is shown. C Hematoxylin and eosin (H&E) and α-RNF4 staining of representative cases of human pan-sarcoma tissue microarray comprising 30 cases; Liposarcoma #884; Osteosarcoma #20886. D H&E and α-BMP6 staining of representative cases of human pan-sarcoma tissue microarray comprising 30 cases. In each set, normal non-tumor tissue is shown. Samples #11308 and #15383 are both leiomyosarcoma. E – G Kaplan–Meier plots of mixed osteosarcoma overall patient survival relative to RNF4, BMP6, and RGMB expression. Blue = high expression, Red = low expression. Total sample size n = 88. Data generated with the online R2 visualization tool, Genomics Analysis and Visualization Platform ( https://hgserver1.amc.nl/cgi-bin/r2/main.cgi ). To establish the optimum survival cut-off by statistical testing, classification in high and low expression was performed using the R2 Kaplan Scan function. The expression graph illustrates p value plotted against mRNA expression to determine the best cut-off. Statistical significance, P , was calculated by log-rank test.

Article Snippet: The antibodies used were: anti-RNF4 (Merck, HPA022047) and BMP6 (Merck, HPA062683).

Techniques: Expressing, Derivative Assay, Generated, Immunostaining, Staining, Microarray