Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 3. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different size of heparin oligosaccharides (from fp4 to dp18). (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively, and concentrations of heparin oligosaccharides were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 4. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different chemical modified heparin in solution. (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively, and concentrations of different modified heparin were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 5. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different GAGs in solution. (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively and concentrations of GAGs were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 protects from IL (interleukin)-33–induced endothelial-to-mesenchymal transition (EndMT) and induces sST2 (soluble supression of tumorigenicity 2) expression in control pulmonary arterial endothelial cells (PAECs) in vitro. A , Representative immunofluorescent staining of PAEC for CD31 (endothelial marker), SM22α (smooth muscle protein 22-alpha; mesenchymal marker), and 4′,6-diamidino-2-phenylindole (DAPI; nuclei). Cells were treated with BMP9 (1 ng/mL), IL-33 (100 ng/mL), both, or left untreated (control [CTR]) for 3 days. Bar graphs show CD31 and SM22α intensity quantification (n=3). B , sST2 , ST2L , CTGF , and PAI-1 gene expressions in PAECs after 16-hour stimulation with TGF (transforming growth factor)-β (1 ng/mL), activin A (50 ng/mL), or untreated (CTR; technical replicates [t.n.]=3). C , sST2 , ST2L , ID1 , and ID3 gene expressions in PAECs after 3-hour stimulation with BMP4 (50 ng/mL), BMP6 (50 ng/mL), BMP9 (1 ng/mL), BMP10 (1 ng/mL), or untreated (CTR; t.n.=3). Statistical analysis: 1-way ANOVA with the Tukey post hoc test; * P <0.05, ** P <0.01, and **** P <0.0001. Data are shown as mean±SD.

Article Snippet: Human recombinant BMP4 (314-BP-010/CF), BMP6 (507-BP-020/CF), BMP9 (3209-BP-010/CF), BMP10 (2926-BP-025/CF), activin A (338-AC-010/CF), TGF-b1 (240-B-010/CF), and recombinant human soluble ST2/IL-33R Fc (523-ST-100) were obtained from R&D Systems.

Techniques: Expressing, Control, In Vitro, Staining, Marker

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 protects from IL (interleukin)-33–induced endothelial-to-mesenchymal transition (EndMT) and induces sST2 (soluble supression of tumorigenicity 2) expression in pulmonary arterial endothelial cells (PAECs) from patients with pulmonary arterial hypertension (PAH) in vitro. A , Representative immunofluorescent staining of PAH PAEC for CD31 (endothelial marker), SM22α (smooth muscle protein 22-alpha; mesenchymal marker), and 4′,6-diamidino-2-phenylindole (DAPI). Cells were treated with BMP9 (1 ng/mL), IL-33 (100 ng/mL), both, or left untreated (control [CTR]) for 3 days. Bar graphs show quantification of CD31 and SM22α intensity (technical replicates [t.n.]=3). B , sST2 , ST2L , CTGF , and Pai-1 gene expressions in PAH PAECs s after 16-hour stimulation with TGF (transforming growth factor)-β (1 ng/mL), activin A (50 ng/mL), or untreated (CTR; t.n.=3). C , sST2 , ST2L , ID1 , and ID3 gene expressions in PAH PAECs after 3-hour stimulation with BMP4 (50 ng/mL), BMP6 (50 ng/mL), BMP9 (1 ng/mL), BMP10 (1 ng/mL), or untreated (CTR; t.n.=3). Statistical analysis: 1-way ANOVA with the Tukey post hoc test; P <0.05, * P <0.01, ** P <0.001, and *** P <0.0001. Data are shown as mean±SD.

Article Snippet: Human recombinant BMP4 (314-BP-010/CF), BMP6 (507-BP-020/CF), BMP9 (3209-BP-010/CF), BMP10 (2926-BP-025/CF), activin A (338-AC-010/CF), TGF-b1 (240-B-010/CF), and recombinant human soluble ST2/IL-33R Fc (523-ST-100) were obtained from R&D Systems.

Techniques: Expressing, In Vitro, Staining, Marker, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 2. C-X-C motif chemokine 10 (CXCL10), stanniocalcin-1, and placental growth factor mRNA expression is significantly increased by trophoblast conditioned media (CM). Messenger RNA expression in vascular spheroids was measured by quantitative real-time PCR after stimulation with trophoblast CM for 24 hours (n=4). Data are expressed as mRNA expression relative to an external calibrator. A, CXCL10. B, Stanniocalcin-1. C, Placental growth factor. All data are presented as mean±SEM. *P<0.05.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, RNA Expression, Real-time Polymerase Chain Reaction

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 3. C-X-C motif chemokine 10 (CXCL10) protein is expressed by vascular spheroids stimulated with trophoblast con- ditioned media. A, Vascular spheroid lysates were examined by Western blot analysis for the presence of CXCL10. Tubulin was used as an internal loading control. The image shown is represen- tative of 3 independent experiments. Densitometric analysis of Western blots (n=3) with mean±SEM CXCL10/tubulin presented as a fold-change over control-treated spheroids. *P<0.05. Vascu- lar spheroids treated with control media (B) or trophoblast condi- tioned media (C) were cryosectioned and sections were examined by immunostaining for the presence of CXCL10. Sections were taken through approximately the center of the spheroid. Negative control incubated with rabbit IgG in place of primary antibody is inset. Scale bar,100 µm.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Western Blot, Control, Immunostaining, Negative Control, Incubation

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 4. C-X-C motif chemokine 10 (CXCL10) protein is expressed by first trimester decidua and dissected spiral arteries stimulated with trophoblast conditioned media. CXCL10 (A and C) and α-smooth muscle actin protein (D) expression was examined by immunohis- tochemistry in serially sectioned first trimester decidua (n=5; representative image shown). CXCL10 and α-smooth muscle actin protein colocalized to the same cells (indicated by arrowheads). Trophoblasts (labeled with CK7, B) were present at this stage of remodeling. Negative control (inset) was incubated with nonimmune IgG in place of primary antibody. Scale bar represents 100 µm or 50 µm in zoom. (E) Expression of CXCL10 (green) and (F) VWF, an endothelial cell marker (red), was examined in a dissected spiral artery treated with extravillous trophoblast (EVT) conditioned media (G, merge). EC indicates endothelial cells; and VSM, vascular smooth muscle. Negative control (inset) was incubated with nonimmune IgG in place of primary antibody. Scale bar, 50 µm.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, Labeling, Negative Control, Incubation, Marker

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 5. The role of IFN-γ in C-X-C motif chemokine 10 (CXCL10) expression. A, Recombinant IFN-γ induces CXCL10 expression in vascular spheroids. Control or rhIFN-γ was added to spheroids made of endothelial cell (EC) alone, vascular smooth muscle cell (VSMC) alone, or cocultured EC/VSMC. CXCL10 expression was measured by Western blot analysis (n=4). B, Neutralizing IFN-γ in extravillous trophoblast (EVT) conditioned media (CM) decreased vascular spheroid CXCL10 expression. Control media or EVT CM was incubated with IFN-γ–neutralizing antibody or corresponding IgG control and added to EC/VSMC spheroids. CXCL10 expression was determined by Western blot analysis. *P<0.05 (n=5).

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, Recombinant, Control, Western Blot, Incubation

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 6. The effects of extravillous tro- phoblast (EVT) conditioned media (CM) and C-X-C motif chemokine 10 (CXCL10) on vascular smooth muscle cells (VSMCs). VSMCs were incubated for 72 hours in media containing 0.5% FCS, then a further 72 hours with indicated concentrations of EVT CM and recombi- nant human CXCL10 (n=at least 5 inde- pendent experiments). Expression of (A) α-smooth muscle actin and (B) calponin was examined by Western blot analysis. Time-lapse microscopy was used to analyze the effects of rhCXCL10 (C) and EVT CM (D) on VSMC motility during a 24-hour incubation. Data are displayed as the mean±SEM of a minimum of 3 pooled experiments. *P<0.05; **P<0.01.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Incubation, Expressing, Western Blot, Time-lapse Microscopy