recombinant human emmprin  (R&D Systems)

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

doi: 10.1186/s12964-024-01508-x

Figure Lengend Snippet: Association of tissue factor (TF) and CD147 overexpression with extracellular vesicle (EV) production and invasion potential. The expression of green fluorescent protein (GFP)-fusion CD147 on the plasma membrane of J82 cells was observed using confocal microscopy ( A ). To assess the vesicular export of CD147, GFP-labeled vesicles were derived from J82 cells transfected with the CD147-GFP construct and analyzed using flow cytometry ( B ). The lysates of the EVs were prepared using 0.1% Triton™ X-100. EVs were derived from human bladder urothelial carcinoma J82 cells co-transfected with TF and CD147 plasmids. The EVs were quantified using NTA in the 48-h culture supernatant of J82 cells overexpressing CD147 and TF versus that of the control transfection condition ( C ). Invasion potency was measured by a Transwell assay, with 6–8 images of the cells that had passed through the coated microporous membrane after 24 h per experiment ( D )

Article Snippet: J82 cells were exposed to their own isolated and characterized microvesicle-like EVs, which were pre-incubated with or without the CD147 inhibitor AC-73 (MedChemExpress LLC, NJ), human recombinant TFPI together with factor VII (Sigma-Aldrich, MO), or human recombinant CD147 protein (rCD147, Sino Biological Europe, Germany) for the specified duration.

Techniques: Over Expression, Expressing, Membrane, Confocal Microscopy, Labeling, Derivative Assay, Transfection, Construct, Flow Cytometry, Transwell Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

doi: 10.1186/s12964-024-01508-x

Figure Lengend Snippet: Impact of the combined inhibitors treatment on extracellular vesicle (EV)-induced migration potential. J82 human bladder urothelial carcinoma cells were exposed to 150 µg/ml protein from J82 cell-derived EVs and treated with either 5 µM CD147 inhibitor AC-73 alone ( A ) or a combination of 2.5 µM AC-73 and tissue factor pathway inhibitor (TFPI, 300 ng/ml, B ) for 16 h. Relative migration was quantified via a Transwell assay utilizing 6–8 images of cells that traversed the microporous membrane per experiment

Article Snippet: J82 cells were exposed to their own isolated and characterized microvesicle-like EVs, which were pre-incubated with or without the CD147 inhibitor AC-73 (MedChemExpress LLC, NJ), human recombinant TFPI together with factor VII (Sigma-Aldrich, MO), or human recombinant CD147 protein (rCD147, Sino Biological Europe, Germany) for the specified duration.

Techniques: Migration, Derivative Assay, Transwell Assay, Membrane

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

doi: 10.1186/s12964-024-01508-x

Figure Lengend Snippet: Impact of treatment with the combined inhibitors on extracellular vesicle (EV)-induced matrix metalloproteinases (MMPs) release. J82 human bladder urothelial carcinoma cells were treated with J82 cell-derived EVs or with a pool of isolated EVs from urine samples collected from patients with muscle-invasive bladder cancer ( A ), along with either 2.5 µM or 5 µM of the CD147 inhibitor AC-73, in the absence or presence of tissue factor pathway inhibitor (TFPI, 300 ng/ml, B ) for 24 h. Recombinant CD147 (rCD147) was used as a positive control to induce MMP activity. The induction of MMPs was measured using colorimetric enzymatic methods

Article Snippet: J82 cells were exposed to their own isolated and characterized microvesicle-like EVs, which were pre-incubated with or without the CD147 inhibitor AC-73 (MedChemExpress LLC, NJ), human recombinant TFPI together with factor VII (Sigma-Aldrich, MO), or human recombinant CD147 protein (rCD147, Sino Biological Europe, Germany) for the specified duration.

Techniques: Derivative Assay, Isolation, Recombinant, Positive Control, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

doi: 10.1186/s12964-024-01508-x

Figure Lengend Snippet: Impact of treatment with the combined inhibitors on extracellular vesicle (EV)-induced invasion potential. J82 human bladder urothelial carcinoma cells were exposed to 150 µg/ml protein from J82 cell-derived EVs or with a pool of isolated EVs from urine samples collected from patients with muscle-invasive bladder cancer, and treated with either 5 µM of the CD147 inhibitor AC-73 alone ( A ) or a combination of 2.5 µM AC-73 and tissue factor pathway inhibitor (TFPI, 300 ng/ml, B ) for 24 h. Invasion potency was measured via a Transwell assay utilizing 6–8 images of cells that traversed the microporous membrane per experiment

Article Snippet: J82 cells were exposed to their own isolated and characterized microvesicle-like EVs, which were pre-incubated with or without the CD147 inhibitor AC-73 (MedChemExpress LLC, NJ), human recombinant TFPI together with factor VII (Sigma-Aldrich, MO), or human recombinant CD147 protein (rCD147, Sino Biological Europe, Germany) for the specified duration.

Techniques: Derivative Assay, Isolation, Transwell Assay, Membrane

Journal: Cell Communication and Signaling : CCS

Article Title: Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells

doi: 10.1186/s12964-024-01508-x

Figure Lengend Snippet: Impact of treatment with the combined inhibitors on the extracellular vesicle (EV) binding capability. J82 human bladder urothelial carcinoma cells were exposed in suspension to carboxyfluorescein succinimidyl ester fluorescent dye (CFSE)-labeled J82 cell-derived EVs and treated with either 5 µM CD147 inhibitor AC-73 alone or a combination of 2.5 µM AC-73 and tissue factor pathway inhibitor (TFPI, 300 ng/ml). The cells were analyzed for green fluorescence in the FITC channel using flow cytometry

Article Snippet: J82 cells were exposed to their own isolated and characterized microvesicle-like EVs, which were pre-incubated with or without the CD147 inhibitor AC-73 (MedChemExpress LLC, NJ), human recombinant TFPI together with factor VII (Sigma-Aldrich, MO), or human recombinant CD147 protein (rCD147, Sino Biological Europe, Germany) for the specified duration.

Techniques: Binding Assay, Suspension, Labeling, Derivative Assay, Fluorescence, Flow Cytometry

Journal: The Journal of Physical Chemistry. C, Nanomaterials and Interfaces

Article Title: Texture or Linker? Competitive Patterning of Receptor Assembly toward Ultra-Sensitive Impedimetric Detection of Viral Species at Gold-Nanotextured Titanium Surfaces

doi: 10.1021/acs.jpcc.3c00697

Figure Lengend Snippet: (A) electrochemical cell and (B) AuTi-NTE electrode photographs. Topography of as-prepared AuTi-NTE, visualized by: (C) AFM ( D) FE-SEM. (E–H) functionalization scheme: (E) unmodified AuTi-NTE, (F) MPA adsorption, (G) consecutive functionalization steps, (H) ready-to-use sensor with CD147 anchored. (I) XPS S2p analysis after modification with MPA.

Article Snippet: The biological materials, i.e., the receptor-transmembrane glycoprotein CD147 (972-EMN) (CD147), and the targeted SARS-CoV-2 Spike receptor-binding domain proteins (CHO: 1-534-CV) (RBD), were obtained from R&D Systems.

Techniques: Adsorption, Modification

Journal: The Journal of Physical Chemistry. C, Nanomaterials and Interfaces

Article Title: Texture or Linker? Competitive Patterning of Receptor Assembly toward Ultra-Sensitive Impedimetric Detection of Viral Species at Gold-Nanotextured Titanium Surfaces

doi: 10.1021/acs.jpcc.3c00697

Figure Lengend Snippet: Adsorption of the CD147 receptor on the gold surface with MPA-EDC_NHS functionalization. (A) Geometry of the CD147 receptor on the Au surface in two perpendicular projections—the tilt of the protein is marked black and equal to ca. 60°; (B and C) schematic illustration of 9 CD147 receptors on the flat gold and AuNP surfaces. Green segments indicate the place at the receptor for binding of the viral protein.

Article Snippet: The biological materials, i.e., the receptor-transmembrane glycoprotein CD147 (972-EMN) (CD147), and the targeted SARS-CoV-2 Spike receptor-binding domain proteins (CHO: 1-534-CV) (RBD), were obtained from R&D Systems.

Techniques: Adsorption, Binding Assay

Journal: Sensors and Actuators. B, Chemical

Article Title: Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: Multivariate data analysis of multisine impedimetric response

doi: 10.1016/j.snb.2022.132427

Figure Lengend Snippet: SARS-CoV-2 receptors used for biofunctionalisation of the BDD electrodes in FS-5. Dimensions and atom counts of receptors are calculated from their crystal structures (IgG2B, ACE2) or models (CD147). Isoelectric points and total charge (in |e|) are calculated from their sequences using Protein Calculator v3.4 (protcalc.net).

Article Snippet: Biological materials utilised in this work, i.e., monoclonal mouse IgG2B anti-SARS-CoV-2 Spike RBD antibodies (MAB1054), recombinant SARS-CoV-2 Spike RBD (HEK: 10500-CV, CHO: 10534-CV), recombinant Human ACE2 (933-ZN), and recombinant Human CD147 Fc Chimera (972-EMN) were obtained from R&D Systems.

Techniques:

Journal: Sensors and Actuators. B, Chemical

Article Title: Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: Multivariate data analysis of multisine impedimetric response

doi: 10.1016/j.snb.2022.132427

Figure Lengend Snippet: Molecular dynamic of biofunctionalised BDD interface: A) rmsd and B) radius of gyration of ACE2 (red) and CD147 (blue) on the surface; C) Scheme of the interaction of SARS-CoV-2 envelope protein with assay; D) visualisation of the spike protein from the crystal structure of SARS-CoV-2 S superimposed on the final snapshot of ACE2 on the NTA-functionalised surface; E) visualisation of the spike protein from the crystal structure of the SARS-CoV-2 S-ACE2 complex superimposed on the final snapshot of CD147 on the NTA-functionalised surface;.

Article Snippet: Biological materials utilised in this work, i.e., monoclonal mouse IgG2B anti-SARS-CoV-2 Spike RBD antibodies (MAB1054), recombinant SARS-CoV-2 Spike RBD (HEK: 10500-CV, CHO: 10534-CV), recombinant Human ACE2 (933-ZN), and recombinant Human CD147 Fc Chimera (972-EMN) were obtained from R&D Systems.

Techniques:

Journal: Sensors and Actuators. B, Chemical

Article Title: Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: Multivariate data analysis of multisine impedimetric response

doi: 10.1016/j.snb.2022.132427

Figure Lengend Snippet: To the left, the charge transfer resistance (R CT ), effective electric double-layer capacitance (C eff ), and CPE exponent (α) changes based on pDEIS results of the first cathodic polarisation scan. The BDD electrode was functionalised with one of the receptor proteins (CD147, ACE2, IgG2B). The test was performed in 1 mM K 3 [Fe(CN) 6 ] and 1 mM K 4 [Fe(CN) 6 ] in PBS solution (black line) as well as with SARS-CoV-2 RBD at different concentrations: 0.301, 3.47 and 50.0 ng/mL (coloured lines, darker with increasing concentration). The dashed line represents the corresponding cathodic currents. To the right, the magnified point of interest (POI) polarisation ranges, characterised with the highest differentiation of electric parameters R CT and C eff with SARS-CoV-2 RBD concentration, based on PCA and PLS analyses.

Article Snippet: Biological materials utilised in this work, i.e., monoclonal mouse IgG2B anti-SARS-CoV-2 Spike RBD antibodies (MAB1054), recombinant SARS-CoV-2 Spike RBD (HEK: 10500-CV, CHO: 10534-CV), recombinant Human ACE2 (933-ZN), and recombinant Human CD147 Fc Chimera (972-EMN) were obtained from R&D Systems.

Techniques: Concentration Assay