human αvβ3 ectodomain  (R&D Systems)

Journal: PLOS ONE

Article Title: An intravenous pancreatic cancer therapeutic: Characterization of CRISPR/Cas9n-modified Clostridium novyi -Non Toxic

doi: 10.1371/journal.pone.0289183

Figure Lengend Snippet: (A) Pictural description of the α V β 3 adhesion assay methodology. (B) Fold change observed in crystal violet (CV) absorbance at a wavelength of 590 for wild type (WT), non-toxic (NT) C . novyi as well as the putative RGD-modified candidates (A and B) after exposure to the α V β 3 coated surface of the adhesion assay. * denotes a p value of < 0.05 when compared to any other cohort, including WT, NT, and modification Candidate B. (C) Average CV pixel count of entire integrin coated surface for candidates A and B as well as wild-type (WT) and non-toxic (NT) C . novyi that remain on the α V β 3 coated surface. ‡ denotes a p value of 0.05 when compared to any other cohort, including WT, NT, and modification Candidate B. Error bars represent standard deviation from the cumulative mean of three experimental replications (n = 6 each) for a total n = 18.

Article Snippet: Purified α V β 3 integrin (100μL of 10μg/mL carrier-free, human recombinant protein, R&D Systems Bio-Techne 3050-AV) solution was administered to the center of the corona-treated cover slips.

Techniques: Cell Adhesion Assay, Modification, Standard Deviation

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Cytotoxic effect and NO assay of integrin α v β 3 treated with or without IL-1β (10 ng/mL) and treated with integrin α v β 3 (0, 0.5, 1, 2, 5, 10, 25, 50, and 100 ng/mL) on Chon-001 cells at 37 °C for 24 h. ( A ) Protein structure of integrin α v β 3 (RCSB PDB ( https://www.rcsb.org , accessed on 15 June 2023), ID 1JV2). ( B ) Integrin α v β 3 (0, 0.5, 1, 2, 5, 10, 25, 50, and 100 ng/mL) was treated by concentration without IL-1β, and further toxicity of integrin α v β 3 to cell for 24 h was measured. ( C ) Integrin α v β 3 treated with (0, 0.5, 1, 2, 5, 10, 25, 50, and 100 ng/mL) concentrations with IL-1β. ( D ) Effect of integrin α v β 3 on IL-1β-induced NO release from Chon-001 cells. Cell viability when IL-1β and integrin α v β 3 were treated together for 24 h. ### p < 0.001 vs. untreated group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs IL-1β treated group.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Concentration Assay

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: The anti-inflammatory effect with or without integrin α v β 3 in IL-1β-induced Chon-001 cells. The relative density of COX-2 and iNOS expression decreased in a dose-dependent manner. ( A ) The expression of COX2 and iNOS were quantified via Western blot analysis. ( B ) Expression of COX2 with or without IL-1β (10 ng/mL). ( C ) Expression of iNOS with or without IL-1β (10 ng/mL). ( D ) Effect of integrin α v β 3 on IL-1β-induced PGE2 release from Chon-001 cells. Comparison with only IL-1β ** p < 0.01, *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group ## p < 0.01, ### p < 0.001.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced MAPKs protein expression in Chon-001 cells. ( A ) The expression of MAPKs was quantified via Western blot analysis. ( B ) Expression of p-ERK affected by integrin α v β 3 . ( C ) Expression of p-JNK affected by integrin α v β 3 . ( D ) Expression of p-p38 affected by integrin α v β 3 . Comparison with only IL-1β * p < 0.05, *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group ## p < 0.01, ### p < 0.001.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced protein expression of p-p65 and p-IκBα in Chon-001 cells. Chon-001 cells were treated with integrin α v β 3 (0, 25, and 50 ng/mL) with or without IL-1β (10 ng/mL) at 37 °C 24 h. ( A ) The expression of NF-κB was quantified via Western blot analysis. ( B ) Relative density of p-IκBα for IκBα. ( C ) Relative density of p-P65 for P65. Comparison with only IL-1β ** p < 0.01, *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group # p < 0.05.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced protein expression of p-STAT3 in Chon-001 cells. ( A ) The expression of p-STAT3 was quantified via Western blot analysis. ( B ) Relative density of p-STAT3 for β-actin. Comparison with only IL-1β *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group ### p < 0.001.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced protein expression of ALP and RUNX2 in Chon-001 cells. ( A ) The expression of ALP and RUNX2 were quantified via Western blot analysis. ( B ) Relative density of ALP for β-actin. ( C ) Relative density of RUNX2 for β-actin. Comparison with only IL-1β *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group ### p < 0.001.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced protein expression of BMP2 and BMP4 in Chon-001 cells. ( A ) The expression of BMP2, BMP4, SOX9, COL1A1, COL2A1, and Aggrecan were quantified via Western blot analysis. ( B ) Relative density of BMP2 for β-actin. ( C ) Relative density of BMP4 for β-actin. ( D ) Relative density of SOX9 for β-actin. ( E ) Relative density of Aggrecan for β-actin. ( F ) Relative density of COL1A1 for β-actin. ( G ) Relative density of COL2A1 for β-actin. Comparison with only IL-1β * p < 0.05, ** p < 0.01, *** p < 0.001. Comparison with integrin α v β 3 and IL-1β-treated group ## p < 0.01, ### p < 0.001.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Effect of integrin α v β 3 with or without IL-1β-induced protein expression of MMP2 and MMP9 in Chon-001 cells. ( A ) The expression of MMP2 and MMP9 was quantified via Western blot. ( B ) A significant decrease in MMP2 from 25 and 50 ng/mL. ( C ) In MMP9, there was a similar protein expression level. Comparison with only IL-1β * p < 0.05, ** p < 0.01. Comparison with integrin α v β 3 and IL-1β-treated group ## p < 0.01.

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques: Expressing, Western Blot, Comparison

Journal: Biomedicines

Article Title: Potential Joint Protective and Anti-Inflammatory Effects of Integrin α v β 3 in IL-1β-Treated Chondrocytes Cells

doi: 10.3390/biomedicines11102745

Figure Lengend Snippet: Schematic representation of anti-inflammation, osteogenesis, and chondrogenesis by integrin α v β 3 .

Article Snippet: Integrin α v β 3 was purchased from biotechne (Minneapolis, MN, USA)—Cat. no. 3050-AV-050.

Techniques:

Journal: The Journal of biological chemistry

Article Title: Receptor-binding domain of SARS-CoV-2 is a functional αv-integrin agonist.

doi: 10.1016/j.jbc.2023.102922

Figure Lengend Snippet: Figure 2. FN-null MEF adhesion to S1-RBD is mediated by αvβ3 integrins and RGD. FN-null MEFs (5 × 105 cells/ml) were preincubated for 30 min with 50 μg/ml integrin-blocking antibodies (A–C) or 25 μM peptide (D–E) before seeding (9.4 × 104 cells/cm2) onto plates precoated with 250 nM S1-RBD (A and D), FNIII10 (B and E), or type I collagen (C). Relative cell number was determined by crystal violet staining. Data are mean ± SEM, normalized to corresponding vehicle (PBS) controls; n = 3 independent experiments performed in triplicate. One-way ANOVA, Bonferroni’s post hoc test: A–C *p < 0.05 versus PBS, IgG; +p < 0.05 versus PBS, IgM; #p < 0.05 versus PBS, anti-α5+β1; D–E *p < 0.05 versus PBS; #p < 0.05 versus corresponding negative control, RAD or KGD.

Article Snippet: Recombinant human integrins αvβ3 (3050-AV), αvβ6 (3817- AV), and α5β1 (3230-A5) were from R&D Systems.

Techniques: Blocking Assay, Staining, Negative Control

Journal: The Journal of biological chemistry

Article Title: Receptor-binding domain of SARS-CoV-2 is a functional αv-integrin agonist.

doi: 10.1016/j.jbc.2023.102922

Figure Lengend Snippet: Figure 3. Recombinant human integrins αvβ3 and αvβ6, but not α5β1, bind to immobilized S1-RBD. Representative kinetic data for αvβ3 (A and D), αvβ6 (B and E), or α5β1 (C and F) binding to immobilized S1-RBD (A–C), FNIII10 (D–E), or FNIII8-10 (F). Data are presented as representative traces (bold colored lines) collected from one of two (α5β1, 100–1000 nM) or three (αvβ3, 5–450 nM and αvβ6, 5–500 nM) experiments and corresponding 1:1 binding fits (black lines).

Article Snippet: Recombinant human integrins αvβ3 (3050-AV), αvβ6 (3817- AV), and α5β1 (3230-A5) were from R&D Systems.

Techniques: Recombinant, Binding Assay

Journal: Nature Medicine

Article Title: Angiopoietin-like 3-derivative LNA043 for cartilage regeneration in osteoarthritis: a randomized phase 1 trial

doi: 10.1038/s41591-022-02059-9

Figure Lengend Snippet: a , LNA043-induced DKK1 secretion is integrin α 5 -dependent. Integrins α 5 and α v were knocked down in C-28/I2 cells by siRNA (siITGA5 and siITGAV, respectively; siITGA5+V for both combined). Left, DKK1 secretion induced by LNA043 (200 μg ml −1 ), as analyzed by ELISA. The data represent means ± s.e.m. of 3–8 independent experiments (from left to right, *** P = 0.0003, * P = 0.0391, * P = 0.0178 and **** P ≤ 0.0001). Statistical significance was determined by one-way ANOVA with mixed-effect analysis and Šídák’s multiple comparison test. Middle, efficiency of the knockdown, as monitored by immunoblotting. GAPDH was used as a loading control. Right, quantification of the immune bands, as determined by densitometry. The data are presented as a percentage decrease ± s.e.m. versus control siRNA (siCtrl) and represent six independent experiments (**** P ≤ 0.0001). Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparison test. b , LNA043 interacts with α 5 β 1 integrin in vitro, as shown by co-IP experimentation. Recombinant human α 5 β 1 integrin (rhα 5 β 1 ) or recombinant human α v β 3 integrin (rhα v β 3 ) were incubated with LNA043 for 1 h at 37 °C. Immunoprecipitation was performed using a monoclonal antibody to α 5 β 1 (anti-α 5 β 1 ) or α v β 3 integrin (anti-α v β 3 ) followed by western blot analysis (right) with monoclonal antibodies to the α 5 (left) or α v (middle) integrin subunit or ANGPTL3, which recognize LNA043 (right). One representative experiment is shown out of two to four independent experiments. c , SPR single-cycle binding studies. LNA043 (top left and top right), FN1 (bottom left) or VTN (bottom right) were immobilized on a CM5 sensor. Integrins α 5 β 1 (left) or α v β 3 (right) served as analytes (0.6–10 µM). The sensorgrams were fitted with a 1:1 binding fit. One representative experiment is shown out of three independent experiments with comparable results. RU, resonance units.

Article Snippet: A total of 200–1,000 ng samples of recombinant human α 5 β integrin (rhα 5 β ; R&D Systems) or recombinant human α v β 3 integrin (rhα v β 3 ; R&D Systems) were incubated with 200 ng LNA043 in DMEM/F-12 medium (Life Technologies) for 1 h at 37 °C.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Knockdown, Western Blot, Control, In Vitro, Co-Immunoprecipitation Assay, Recombinant, Incubation, Immunoprecipitation, Bioprocessing, Binding Assay

Journal: Nature Medicine

Article Title: Angiopoietin-like 3-derivative LNA043 for cartilage regeneration in osteoarthritis: a randomized phase 1 trial

doi: 10.1038/s41591-022-02059-9

Figure Lengend Snippet: LNA043 binding to the FN1 receptor, integrin α 5 β 1 , and intracellular signaling e.g. via FAK and AKT phosphorylation induces transcriptional counter-regulation of OA-relevant gene expression. Specifically, a, up-regulation of essential cartilage matrix components PRG4 , COL2A1 , ACAN , COMP , MATN4 , COL9A1 results in cartilage matrix synthesis; b, regulation of genes DKK1 , FRZB , WNT16 inhibits WNT signaling, which induces chondrogenesis and cartilage regeneration; c, regulation of genes CHRDL2 , GREM1 , BMP6 inhibits BMP signaling, which prevents detrimental chondrocyte hypertrophy and mineralization in OA; d, downregulation of OA progression mediators OPN , DNER , OPG as well as FN 1 splice variants and FN1 , which in the OA environment is cleaved into cartilage degradation-promoting fragments, reduces cartilage degeneration and progression of OA. Transparent symbols depict transcriptomics results, colored symbols in vitro assay-confirmed data.

Article Snippet: A total of 200–1,000 ng samples of recombinant human α 5 β integrin (rhα 5 β ; R&D Systems) or recombinant human α v β 3 integrin (rhα v β 3 ; R&D Systems) were incubated with 200 ng LNA043 in DMEM/F-12 medium (Life Technologies) for 1 h at 37 °C.

Techniques: Binding Assay, Phospho-proteomics, Gene Expression, In Vitro