Journal: PLoS ONE

Article Title: Inhibition of Cellular Adhesion by Immunological Targeting of Osteopontin Neoepitopes Generated through Matrix Metalloproteinase and Thrombin Cleavage

doi: 10.1371/journal.pone.0148333

Figure Lengend Snippet: (A) Cellular adhesion in percent of HEK 293 cells from 0 to 30 nM coated recombinant OPN forms in 3 nM steps. Depicted are the means ± SEM of 3 independent experiments. * and + indicate significance of cellular adhesion to mOPN or tOPN in comparison to full length OPN, respectively. (B) Blockade of cellular adhesion of HEK 293 cells at 10 or 30 nM coated recombinant OPN forms with 1μM antagonistic integrin inhibitors. RGES (black bars) was used as a control peptide. Cilengitide (white bars) inhibits the integrins α V β 3 , α V β 5 , and α 5 β 1 . TR-14035 (hatched bars) inhibits the integrins α 4 β 7 and α 4 β 1. Depicted are the means ± SEM of 3 independent experiments. * indicate significance of cell adhesion to the RGES control peptide.

Article Snippet: For integrin inhibition the assays were performed by incubating the cells with 1 μM RGES peptide (Abbiotec, San Diego, CA, USA), cilengitide, or TF-14035 (both Medchemexpress, Princeton, NJ, USA) during the 30 minutes labeling and the 30 minutes incubation on the plates.

Techniques: Recombinant, Comparison, Control

Journal: Science Advances

Article Title: Tumor cell–organized fibronectin maintenance of a dormant breast cancer population

doi: 10.1126/sciadv.aaz4157

Figure Lengend Snippet: ( A ) Experimental timeline of inhibitor dosing. Day 7 experiments were dosed continually through a day 7 endpoint; day 28 experiments were dosed continually through a day 28 endpoint; separate cultures were established for 21 days, where inhibitor dosing was initiated for an additional 7 days (dose-established culture). ( B ) HCC 1954 survival at day 7 on collagen coverslips with selected inhibitors dosed for the duration of the experiment. ( C ) Survival of HCC 1954 cells after establishment of dormant culture for 21 days and then subsequent dosing with inhibitors through days 21 to 28. ( D ) HCC 1954 survival at day 7 with inhibitors when seeded onto HCC 1954 decellularized ECM. Black, control; blue, anti-β 1 (MAB17781; 1 μg/ml); green, FAK inhibitor 14 (10 μM); purple, PD0325901 (MEK inhibitor, 10 μM); red, FR180204 (ERK inhibitor, 20 μM); gray, cilengitide (100 μM). P < 0.05 is denoted with *, P ≤ 0.001 with *** and P ≤ 0.0001 with ****.

Article Snippet: Where noted, inhibitors or antibodies were included in the serum-free or serum-containing medium at the following concentrations: E64 (Tocris; 5 μM), ERK inhibitor FR180204 (Sigma-Aldrich; 20 μM), FAK inhibitor 14 (Sigma-Aldrich; 10 μM), soluble fibronectin (Millipore; 10 μg/ml), anti-α5 AF1864 (R&D Systems; 1 μg/ml), cilengitide (Apex Biotechnology; 100 μM), anti-β1 MAB17781 (R&D Systems; 1 μg/ml), MEK inhibitor PD0325901 (Sigma-Aldrich; 10 μM), pan-MMP inhibitor GM6001 (Millipore; 25 μM), ROCK inhibitor Y-27632, (Millipore; 0.1, 1, and 10 μM), pan-TGFβ AB-100-NA (R&D Systems; 20 μg/ml), anti-TGFBR1, LY-364947 (Sigma-Aldrich; 5 μM).

Techniques: Control

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Integrin αvβ3 Induces HSP90 Inhibitor Resistance via FAK Activation in KRAS -Mutant Non–Small Cell Lung Cancer

doi: 10.4143/crt.2021.651

Figure Lengend Snippet: A FAK inhibitor, and not an integrin inhibitor, shows synergistic effects with AUY922 in AUY922-resistant cells. (A, B) Synergistic effect of a cilengitide and AUY922 combination. Cell viability assays were performed in A549R and H1944R cells treated with 1,000 nM of AUY922 plus increasing concentrations of cilengitide for 3 days. (C, D) Synergistic effect of a TAE226 and AUY922 combination. Cell viability assays were performed in A549R and H1944R cells treated with 1,000 nM of AUY922 plus increasing concentrations of TAE226 for 3 days. The confidence interval (CI) was calculated using CalcuSyn software. (E) Western blot analysis of pFAK in A549R cells after each combination treatment. (F) Twelve mice harboring subcutaneous xenograft tumors derived from A549/ITGAvB3 cells were randomized into four groups. After the tumor volume reached 100 mm 3 , AUY922 at 10 mg/kg was administered 3 days/wk, and TAE226 (25 mg/kg) was administered 5 days/wk for up to 21 days. The tumor size was assessed at least three times a week. (G) Western blotting analysis detected cleaved PARP in the mouse xenograft tumors. CI, combi nation indexes; Fa, fraction affected; FAK, focal adhesion kinase; PARP, poly(ADP-ribose) polymerase; SD, standard deviation.

Article Snippet: The human KRAS -mutated NSCLC cell lines, H23, H358, H647, H1944, and A549, were purchased from the American Type Culture Collection (Manassas, VA) and grown at 37°C in 5% CO 2 using RPMI-1640 (Welgene Inc., Gyeonsan, Korea) containing 10% fetal bovine serum (FBS) and 1x penicillin-streptomycin solutions from Welgene Inc. AUY922 (HSP90 inhibitor, luminespib), cilengitide trifluoroacetate (inhibitor of the αvβ3 receptor), and TAE226 (focal adhesion kinase [FAK] inhibitor) were purchased from Selleck Chemicals (Houston, TX).

Techniques: Software, Western Blot, Derivative Assay, Standard Deviation

Journal: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 1. Binding assays. Competition binding of Alexa488-2.5F monomer to integrin receptors expressed on U87MG cells (A) and MDA-MB-231 cells (B) by knottin monomer, dimers, and Cilengitide, as measured by

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques: Binding Assay

Journal: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 2. Effect of D2.5F_1 dimer (red), 2.5F monomer (blue), or Cilengitide (green) on U87MG cell migration compared to untreated control (brown). (A) Bright field images showing tumor cell migration into

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques: Migration, Control

Journal: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 3. Effect of D2.5F_1 dimer (red), 2.5F monomer (blue), or Cilengitide (green) on MDA-MB-231 cell

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques: