bmp3  (Novus Biologicals)

Journal: Nature Communications

Article Title: Role of PDGFRA + cells and a CD55 + PDGFRA Lo fraction in the gastric mesenchymal niche

doi: 10.1038/s41467-023-43619-y

Figure Lengend Snippet: a In isolation, corpus and antral SEMFs are distinct and antral SEMFs divide further into two cell populations. Right: Fgf7 , Nrg1 , Bmp3 , and Ctgf mRNA densities projected on the UMAP plot. b Relative expression of scRNA-seq markers from the three SEMF populations. Circle diameters and fill colors represent the fraction of cells expressing a gene and normalized average expression levels. The two antral SEMF populations differ in expression of Fgf7, Nrg1 , Bmp3 , and other factors; corpus SEMFs resemble the smaller antral Bmp3 + cluster more than the larger population of antral Fgf7 + Nrg1 + cells. c Fluorescence in situ hybridization (RNAscope) on antral tissue sections localizes Fgf7 - and Nrg1 -expressing PDGFRA Hi SEMFs near gland pits and Bmp3 - and Ctgf -expressing cells in the lower half of glands. Images represent fields examined in three independent experiments with each probe. Scale bars 50 μm. Heatmap: average fluorescence signal quantified along 16–25 individual glands ( Ctgf probe quantification is shown in Fig. ). Source data are provided as a Source Data file. Right: schematic representation of antral SEMF distribution. d Co-culture of antral and corpus glands with unfractionated PDGFRA Hi cells, which fail to induce spheroids in the absence of rNOG and RSPO1. Glands cultured in ENRWG medium serve as controls. Scale bars 400 μm. Bars represent mean ± SEM. Antrum: +ENRWG n = 22, +PDGFRA Hi n = 19, No cells n = 22; Corpus: +ENRWG n = 19, +PDGFRA Hi n = 12, No cells n = 18. n : number of culture wells analyzed over six independent experiments. Significance of differences determined by one-way ANOVA coupled with Sidak’s multiple comparison test. **** p < 0.0001, ns not significant. Source data are provided as a Source Data file. e Antral glands exposed to recombinant rFGF7 or rFGF10 in addition to complete ENRWG medium. Budding of spheroid structures increased upon treatment with rFGF7 without affecting spheroid numbers ( n = 8 independent experiments). Bars represent mean ± SEM. Significance of differences determined by one-way ANOVA coupled with Dunnett’s multiple comparison test. ns not significant. Scale bars 400 μm. Source data are provided as a Source Data file.

Article Snippet: mRNA detection for spatial localization of cell populations was carried out using the RNAscope Multiplex Fluorescent Reagent Kit v2 (Advanced Cell Diagnostics) , . and probes designed by Advanced Cell Diagnostics: Cd55 (421251), Grem1 (314741), Ctgf (314541), Fgf7 (443521), Bmp3 (428461), Nrg1 (468841), and Rspo3 (483781).

Techniques: Isolation, Expressing, Fluorescence, In Situ Hybridization, RNAscope, Co-Culture Assay, Cell Culture, Comparison, Recombinant

Journal: Nature Communications

Article Title: Role of PDGFRA + cells and a CD55 + PDGFRA Lo fraction in the gastric mesenchymal niche

doi: 10.1038/s41467-023-43619-y

Figure Lengend Snippet: Antral in situ hybridization localized ( a ) Cd55 to the epithelium (white arrows) and PDGFRA Lo mesenchymal cells (orange arrows) near gland bottoms; ( b ) Grem1 and Ctgf near the gland base (dotted lines demarcate glands, heatmaps: fluorescence along individual glands, n = 35 to n = 39, Source data provided as a Source Data file); and ( c ) Grem1 in SMA1 + muscularis mucosae, extending into sub-glandular PDGFRA Lo cells (dashed lines demarcate glands). Dashed boxes are magnified below ( a ) or to the right ( b , c ). Images represent fields examined in 2 ( a , c ) or 3 ( b ) independent experiments. Scale bars 50 μm. d Schema of mesenchymal cell distributions. MM muscularis mucosae. e Co-culture of gastric glands with CD55 + or CD55 − PDGFRA Lo cells from each region. ENRWG corpus n = 4, antral n = 12; CD55 + corpus n = 4, antral n = 11; CD55 − corpus n = 5, antral n = 9; No cells corpus n = 4, antral n = 11. Bars: mean ± SEM. Significance: one-way ANOVA and Tukey’s multiple comparison test. ns not significant. Source data provided as a Source Data file. Scale bars 400 μm. f Spheroid formation from SI crypts and antral or corpus glands co-cultured with CD55 + (gastric) or CD81 + (SI trophocyte) fractions from each region. SI crypts: CD81 + n = 6, Antral CD55 + n = 5, Corpus CD55 + n = 3; Antral glands: SI CD81 + n = 12, Antral CD55 + n = 11, Corpus CD55 + n = 3; Corpus glands: SI CD81 + n = 4, Antral CD55 + n = 3, Corpus CD55 + n = 4. n : independent experiments, each with ≥2 technical replicates. Bars: mean ± SEM. Significance: one-way ANOVA and Tukey’s multiple comparison test. ns not significant. Source data provided as a Source Data file. g Co-culture of gastric glands with or without corresponding CD55 + PDGFRA Lo cells, growth factors (+GF or −GF), and Porcupine inhibitor Wnt-C59 (Pi) or BMP2/4/7 cocktail (Bs). Bars: mean ± SEM. No cells+GF corpus n = 16, antral n = 24; −CD55 + +Pi corpus and antral n = 9, −CD55 + +Bs corpus n = 8, antral n = 7; −GF corpus n = 18, antral n = 25; +CD55 + corpus n = 13, antral n = 18; +CD55 + +Pi corpus n = 12, antral n = 8; +CD55 + +Bs corpus n = 13, antral n = 9. Significance: one-way ANOVA and Sidak’s multiple comparison test. ns not significant. Source data provided as a Source Data file. Scale bars 400 μm.

Article Snippet: mRNA detection for spatial localization of cell populations was carried out using the RNAscope Multiplex Fluorescent Reagent Kit v2 (Advanced Cell Diagnostics) , . and probes designed by Advanced Cell Diagnostics: Cd55 (421251), Grem1 (314741), Ctgf (314541), Fgf7 (443521), Bmp3 (428461), Nrg1 (468841), and Rspo3 (483781).

Techniques: In Situ Hybridization, Fluorescence, Co-Culture Assay, Comparison, Cell Culture

Journal: Nature Communications

Article Title: Role of PDGFRA + cells and a CD55 + PDGFRA Lo fraction in the gastric mesenchymal niche

doi: 10.1038/s41467-023-43619-y

Figure Lengend Snippet: a In isolation, gastric PDGFRA + mesenchyme other than SEMFs resolves into five subpopulations: previously identified AntInt, AntLo and CorpLo1 (Fig. ) and two CorpLo subpopulations, CorpLo2 and CorpLo3. Relative expression of selected scRNA-seq markers is plotted below. Circle diameters: cell fraction expressing a gene, fill shades: normalized average expression. CorpLo3 shares markers identified in AntInt (see Fig. ). b Bmp5, Bmp7, Wnt4, Grem1, Rspo3 , and Ctgf transcript densities projected on the UMAP plot of resolved non-SEMF cell clusters. CorpLo3 and AntInt express BMPs and Wnt4 , while AntLo, Corp1, and Corp2 express Grem1, Rspo3 , and Ctgf . Dashed lines mark the distinct populations. c Left: Relative Cd55 and Cd34 expression in sub-epithelial cell fractions from scRNA-seq analysis. Circle diameters and fill colors represent the parameters defined in ( a ). Middle: Relative Cd55 expression in PDGFRA Lo subpopulations. Right: Projection of Cd55 density on the PDGFRA Lo UMAP, showing nearly exclusive expression in CorpLo1, CorpLo2 and AntLo cells. d In situ hybridization (RNAscope) of corpus tissue sections, together with LAM immunostaining, localizes Cd55 in the epithelium (white arrows) and in PDGFRA Lo mesenchymal cells (orange arrows) near the gland base, on both sides of muscularis mucosae. The area within the dashed box is magnified below. Images represent scores of fields examined in two independent experiments. Scale bars 50 μm. e In situ hybridization (RNAscope) of corpus tissue sections localizes Grem1 (left) and Rspo3 (middle) near the gland base. Images represent scores of fields examined in two independent experiments. Scale bars 50 μm. Dotted lines mark the muscularis mucosae. In the heatmap, fluorescence signal strength is quantified along 50–52 individual glands. Source data are provided as a Source Data file. f In situ hybridization (RNAscope) of corpus tissue localizes Grem1 in the SMA1-immunostained muscularis mucosae, demarcated by dotted lines, and in the space corresponding to PDGFRA Lo cells near the gland base ( n = 2 independent experiments). Scale bars 50 μm. g Schematic illustration of the distribution of CD55 + PDGFRA Lo cells relative to corpus glands and the muscularis mucosae (MM). Grem1 - and Rspo3 -expressing cells lie largely beneath glands.

Article Snippet: mRNA detection for spatial localization of cell populations was carried out using the RNAscope Multiplex Fluorescent Reagent Kit v2 (Advanced Cell Diagnostics) , . and probes designed by Advanced Cell Diagnostics: Cd55 (421251), Grem1 (314741), Ctgf (314541), Fgf7 (443521), Bmp3 (428461), Nrg1 (468841), and Rspo3 (483781).

Techniques: Isolation, Expressing, In Situ Hybridization, RNAscope, Immunostaining, Fluorescence

Journal: World Journal of Gastroenterology

Article Title: Emerging role of non-invasive and liquid biopsy biomarkers in pancreatic cancer

doi: 10.3748/wjg.v29.i15.2241

Figure Lengend Snippet: Cell free protein (secreted proteome) involved in pancreatic cancer diagnostic and prognostics

Article Snippet: BMP3 , Up , Plasma , TGFβ , Cell signaling , P.

Techniques: Diagnostic Assay, Clinical Proteomics, Methylation, Mutagenesis, Ubiquitin Proteomics, Transduction, DNA Methylation Assay