α 5 β 1 proteins  (R&D Systems)

Journal: Stem Cells International

Article Title: Disrupted Endothelial Cell Layer and Exposed Extracellular Matrix Proteins Promote Capture of Late Outgrowth Endothelial Progenitor Cells

doi: 10.1155/2016/1406304

Figure Lengend Snippet: The influence of fibronectin on the dynamic interaction of LO-EPC with control HAEC and HAEC with ischaemia reperfusion injury under 0.7 dyn/cm 2 shear stress. 3 × 10 4 HAEC were plated into Ibidi μ -Slide VI 0.4 Luer slides which had been precoated with 100 μ g/mL fibronectin. The data represent a mean ± SE of three experiments. LO-EPC adhesion to fibronectin was integrin dependent (b). LO-EPC binding to fibronectin was blocked by antibodies against integrin α 5 β 1, integrin α v β 1, and integrin α v β 3. The data represent the mean ± SE of three experiments. ∗∗ p < 0.01 compared to the control (no blocking antibodies).

Article Snippet: 1.5 mL of 4 × 10 5 LO-EPC was incubated with 10 μ g/mL anti-ITG α 5 β 1 (Millipore, UK), anti-ITG α V β 3 (Millipore, UK), and anti-ITG α v β 1 (Bioss, Antibodies-online.com) or no antibodies (control) for 30 minutes at 37°C with gentle rotation.

Techniques: Binding Assay, Blocking Assay

Journal: Contrast Media & Molecular Imaging

Article Title: Synthesis and Bioevaluation of Iodine-131 Directly Labeled Cyclic RGD-PEGylated Gold Nanorods for Tumor-Targeted Imaging

doi: 10.1155/2017/6081724

Figure Lengend Snippet: (a) Immunofluorescence staining of integrin α v β 3 in B16F10 (upper row) and MCF-7 (lower row) cells. The nucleus were counterstained with DAPI. The red fluorescence intensity is proportional to the expression level of integrin α v β 3 (×200). (b) Results of cell binding assays at various time points.

Article Snippet: The expression of integrin α v β 3 was confirmed by immunofluorescence with a primary anti-integrin α v β 3 antibody (1 : 100, Bioss, Beijing, China) and Cy3-conjugated goat anti-rabbit secondary antibody (1 : 50, Aspen, Wuhan, China) as described previously [ ].

Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, Binding Assay

Journal: Contrast Media & Molecular Imaging

Article Title: Synthesis and Bioevaluation of Iodine-131 Directly Labeled Cyclic RGD-PEGylated Gold Nanorods for Tumor-Targeted Imaging

doi: 10.1155/2017/6081724

Figure Lengend Snippet: (a) Autoradiography of tumor tissue and organs. B16F10 tumor-bearing mice (upper row) and MCF-7 tumor-bearing mice (lower row). (b) Immunofluorescence staining of integrin α v β 3 in B16F10 (upper row) and MCF-7 (lower row) tumors (×200).

Article Snippet: The expression of integrin α v β 3 was confirmed by immunofluorescence with a primary anti-integrin α v β 3 antibody (1 : 100, Bioss, Beijing, China) and Cy3-conjugated goat anti-rabbit secondary antibody (1 : 50, Aspen, Wuhan, China) as described previously [ ].

Techniques: Autoradiography, Immunofluorescence, Staining

Journal: Bone Research

Article Title: Ciliary parathyroid hormone signaling activates transforming growth factor-β to maintain intervertebral disc homeostasis during aging

doi: 10.1038/s41413-018-0022-y

Figure Lengend Snippet: PTH induces integrin α v β 6 expression to activate latent TGF-β. a Immunostaining images showing various types of integrin expressions in IVD tissue from 18-month-ld mice injected with PTH or vehicle and quantitative analysis ( b ). Scale bar, 50 μm. c qRT-PCR analysis of the mRNA levels of various integrin in NP tissue from 18-month-old mice injected with PTH or vehicle. Results reported as fold change. d Western blot analysis of integrin β 6 expression in NP cells of 18-month-old mice at different time points post PTH injection (PTH1-34, 100 nmol·L -1 ). e , f Chromatin immunoprecipitation assay with four different potential pCREB binding sites (primers 1, 2, 3 and 4) in the β6 integrin promoter. g pCREB, Integrin α V β 6 , pSmad2/3, or Safranin-O staining of IVD sections from an IVD ex vivo compression model of 30-month-old rat with treatment of either vehicle or PTH (PTH1-34, 100 nmol·L -1 ). Scale bar, 20 μm. h Quantitative analysis of the percentage of pCREB, pSmad2/3 positive cells and the Integrin α V β 6 positive areas as a percentage of total IVD area (Ar) of ( g ). All data are reported as the mean ± s.d. * P < 0.05. n = 8 per group. Statistical significance was determined by one-way ANOVA and Student's t -test

Article Snippet: Sections for immunostaining were processed using a standard protocol and incubated with primary antibodies to rabbit ACAN (Abcam, 1:100), CCN2 (Abcam, 1:100), integrin β 8 (Abcam, 1:200), pCREB (Abcam, 1:100), and PTH1R PRB-635P (Covance, 1:100), mouse pSmad2/3 (Santa Cruz, 1:100), integrin α v β 6 (Millipore, 1:100), integrin α v β 3 (Bioss, 1:100), integrin α v β 5 (Bioss, 1:100) at 4 °C overnight.

Techniques: Expressing, Immunostaining, Injection, Quantitative RT-PCR, Western Blot, Chromatin Immunoprecipitation, Binding Assay, Staining, Ex Vivo

Journal: ACS Omega

Article Title: Design and Evaluation of Short Self-Assembling Depsipeptides as Bioactive and Biodegradable Hydrogels

doi: 10.1021/acsomega.7b01641

Figure Lengend Snippet: Half inhibition (i.e., IC 50 ) of the fluorescence of FITC-GRGDSP occurs at a ∼40-fold lower concentration for RGD than R-Glc-D, indicating greater affinity of RGD for integrin than R-Glc-D (A). Comparison of FP inhibition by R-Glc-D and RGE peptides (negative control). The average IC 50 value for R-Glc-D is 1115 ± 211 μM (mean ± standard deviation). RGE was tested once as a negative control, and the calculated IC 50 value was 2840 μM (B).

Article Snippet: Human recombinant integrin α 5 β 1 (R&D Systems) was diluted to 900 nM in the same Tris buffer.

Techniques: Inhibition, Fluorescence, Concentration Assay, Comparison, Negative Control, Standard Deviation

Journal: ChemistryOpen

Article Title: Insights into the Binding of Cyclic RGD Peptidomimetics to α 5 β 1 Integrin by using Live‐Cell NMR And Computational Studies

doi: 10.1002/open.201600112

Figure Lengend Snippet: Docking best poses of a) compound 7 (green) and b) compound 6 (green) in the crystal structure of the extracellular domain of α 5 β 1 integrin (α 5 subunit pink, β 1 subunit cyan, model from 3VI4.pdb). Only selected integrin residues involved in interactions with the ligand are shown. Non polar hydrogens are hidden for clarity, whereas intermolecular hydrogen bonds are shown as dashed lines.

Article Snippet: Purified recombinant human integrin α 5 β 1 (R&D Systems, Inc., Minneapolis, MN, USA) was diluted to 0.5 μg mL −1 in coating buffer containing 20 m m Tris‐HCl (pH 7.4), 150 m m NaCl, 1 m m MnCl 2 , 2 m m CaCl 2 , and 1 m m MgCl 2 .

Techniques:

Journal: ChemistryOpen

Article Title: Insights into the Binding of Cyclic RGD Peptidomimetics to α 5 β 1 Integrin by using Live‐Cell NMR And Computational Studies

doi: 10.1002/open.201600112

Figure Lengend Snippet: Docking binding modes: a) A and b) B of compound 3 (green) in the crystal structure of the extracellular domain of α 5 β 1 integrin (α 5 subunit pink, β 1 subunit cyan, model from 3VI4.pdb). Only selected integrin residues involved in interactions with the ligand are shown. Non polar hydrogens are hidden for clarity, whereas intermolecular hydrogen bonds are shown as dashed lines.

Article Snippet: Purified recombinant human integrin α 5 β 1 (R&D Systems, Inc., Minneapolis, MN, USA) was diluted to 0.5 μg mL −1 in coating buffer containing 20 m m Tris‐HCl (pH 7.4), 150 m m NaCl, 1 m m MnCl 2 , 2 m m CaCl 2 , and 1 m m MgCl 2 .

Techniques: Binding Assay

Journal: ChemistryOpen

Article Title: Insights into the Binding of Cyclic RGD Peptidomimetics to α 5 β 1 Integrin by using Live‐Cell NMR And Computational Studies

doi: 10.1002/open.201600112

Figure Lengend Snippet: Inhibition of biotinylated fibronectin binding to α 5 β 1 integrin compared with inhibition of biotinylated vitronectin binding to α v β 3 .

Article Snippet: Purified recombinant human integrin α 5 β 1 (R&D Systems, Inc., Minneapolis, MN, USA) was diluted to 0.5 μg mL −1 in coating buffer containing 20 m m Tris‐HCl (pH 7.4), 150 m m NaCl, 1 m m MnCl 2 , 2 m m CaCl 2 , and 1 m m MgCl 2 .

Techniques: Inhibition, Binding Assay

Journal: British Journal of Cancer

Article Title: Circulating galectins -2, -4 and -8 in cancer patients make important contributions to the increased circulation of several cytokines and chemokines that promote angiogenesis and metastasis

doi: 10.1038/bjc.2013.793

Figure Lengend Snippet: Galectin-induced cytokine secretion enhances expression of the endothelial cell surface adhesion molecules, which are responsible for galectin-mediated cancer cell-endothelial adhesion. ( A ) The presence of galectins induces expressions of cell surface adhesion molecules. Human micro-vascular lung endothelial cells were treated with control 1.5 μ g ml −1 BSA (red colour) or 1.5 μ g ml −1 galectins -2 (purple), -4 (brown), -8 (green), a combination of G-CSF (0.25 ng ml −1 ), IL-6 (0.15 ng ml −1 ), GRO α (1 ng ml −1 ), MCP-1 (1 ng ml −1 ) (black) for 24 h before the expressions of the HMVEC surface integrin α v β 1, VCAM-1, CD44 and E-selectin were analysed by flow cytometry. ( B ) The presence of neutralising antibodies against cell surface adhesion molecules inhibits galectins -2, -4 or -8-mediated cancer cell adhesion. Human micro-vascular lung endothelial cells were treated without or with 1.5 μ g ml −1 galectins -2, -4 or -8 for 24 h. The culture media were collected and used to assess ACA19 − cell adhesion to fresh HMVEC-Ls without or with addition of a combination of neutralising antibodies against integrin α v β 1 (10 μ g ml −1 ), CD44 (10 μ g ml −1 ), VCAM-1(10 μ g ml −1 ) and E-selectin (10 μ g ml −1 ). ** P <0.01, *** P <0.001 (ANOVA, Bonferroni).

Article Snippet: Recombinant human galectins -2, -4 and -8 (residual endotoxin levels <1.0 EU μ g −1 protein), and human Cytokine Protein Array, antibodies against CD44, E-selectin, VCAM-1 and integrin α v β 1 were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Control, Flow Cytometry