Journal: Journal of nanobiotechnology

Article Title: Thrombolytic therapy based on lyophilized platelet-derived nanocarriers for ischemic stroke.

doi: 10.1186/s12951-023-02206-5

Figure Lengend Snippet: Fig. 1 A Schematic representation of the preparation of rtPA-loaded CSM derived from platelets. B Representative mean hydrodynamic diameter of CSM and CSM@rtPA before and after lyophilization (CSM@rtPA/L) process measured by DLS. Data represent mean ± SEM (n = 3, independent samples). C Representative mean diameter of CSM and CSM@rtPA before and after lyophilization process (CSM@rtPA/L) measured by NTA. Data represent mean ± SEM (n = 3, independent samples). D Representative STEM-in SEM images of CSM@rtPA negatively stained with uranyl acetate. Scale bars: 200 nm. E Loading capacity of rtPA encapsulated in CSM samples. Data represent mean ± SEM (n = 3, independent samples). F Representative density plots and quantitative analysis of CSM and CSM@rtPA measured by FC. Scatter density plots of Green fluorescence signal (Green-B channel, rtPA channel) versus Red fluorescence signal (Red-R channel, CellMask DeepRed channel) for CSM, CSM@rtPA, and CSM@rtPA sample after labeling with CellMask Deep Red for lipid staining. Mean fluorescence intensities (MFI) for Green-B channel (D) and Red-R channel. Data represent mean ± SEM (n = 3, independent samples). G. Schematic representation of platelets and platelet-derived CSM surface proteins studied by APC-fluorescently labeled antibodies: anti-hCD47 Ab and anti-hCD42b/GPlba Ab. In vitro Ab binding to platelets and CSM@rtPA before and after lyophilization process. Representative MFI histogram of Red-R channel (APC signal) for platelets, CSM@rtPA and CSM@rtPA/L samples after incubation with APC-anti-CD47 and APC-anti-CD42b/GPIbα antibodies

Article Snippet: Then, fluorescently antibodies, human anti-CD47 APC conjugated Antibody (FAB4670A, R&D Systems) and human antiCD42b/GPlbα APC conjugated Antibody (FAB4067A, R&D systems) were incubated with platelets and with CSMs (10 μL of antibody stock solution /106 cells) and the samples were analyzed by flow cytometer.

Techniques: Derivative Assay, Lyophilization, Staining, Fluorescence, Labeling, In Vitro, Binding Assay, Incubation

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: CD47 levels in human lung adenocarcinoma cells were increased following tumor metastasis (A and B) Comparison of CD47 expression between primary lung adenocarcinoma and lung adenocarcinoma metastases in the lymph nodes. (C and D) Comparison of CD47 expression between primary lung adenocarcinoma and lung adenocarcinoma metastases in the liver. (A) and (C) showed representative IHC images for CD47 staining from 14 to 5 lung adenocarcinoma tissue samples, respectively. Data were presented as means ± SDs. ∗∗p < 0.01. Scale bar, 50 μm.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Comparison, Expressing, Staining

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: Induction of CD47 by IFN-γ in human lung cancer cells (A) Flow cytometry analysis of CD47 surface expression in human lung cancer cell lines upon incubation with recombinant IFN-γ (100 ng/mL, 24 h). Right: Representative image; left: quantitative analysis. (B) Immunofluorescence analysis of CD47 expression in human lung cancer cell lines upon incubation with recombinant IFN-γ (100 ng/mL, 24 h). Scale bar, 20 μm. (C) Western blot analysis of CD47 expression in human lung cancer cell lines after IFN-γ treatment. (D) qRT-PCR analysis of CD47 mRNA in human lung cancer cell lines upon incubation with recombinant IFN-γ (100 ng/mL, 24 h). Left: Representative image; right: quantitative analysis. Data from more than 3 independent experiments were presented as means ± SDs. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Flow Cytometry, Expressing, Incubation, Recombinant, Immunofluorescence, Western Blot, Quantitative RT-PCR

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: Identification of genes involved in the IFN signaling pathway that upregulates CD47 expression (A) qRT-PCR analysis of various genes involved in the IFN signaling pathway in A549 cells upon incubation with recombinant IFN-γ (100 ng/mL, 24 h). (B) Normalized luciferase reporter expression of A549 cells transduced with different sets of lentiviral shRNA constructs. Upper: Schematic representation of the CD47-Prom-Firefly Luciferase-EF1AProm-Renilla luciferase constructs used to generate the reporter cell lines. Lower: Quantitative analysis. (C) IRF1 protein levels in human lung cancer cells upon incubation with recombinant IFN-γ (100 ng/mL, 24 h). Upper: Representative image; lower: quantitative analysis. Data from more than 3 independent experiments were presented as means ± SDs. ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Expressing, Quantitative RT-PCR, Incubation, Recombinant, Luciferase, Transduction, shRNA, Construct

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: IFN-γ induces CD47 expression in lung cancer cells through IRF-1 (A) Sequence of the CD47 promoter showing the position of the putative IRF-1-binding site. (B) Reporter assay for putative IRF-1 binding. The results were normalized to relative luciferase units (RLUs). (C) ChIP assay analyzing the CD47 promoter in A549 cells. The results were normalized to the input. (D and E) IRF-1 mRNA (D) and protein (E) levels in A549 cells transfected with IRF-1-specific shRNA or scramble shRNA after IFN-γ treatment. (F and G) CD47 level in A549 cells transfected with IRF-1-specific shRNA or scramble shRNA after IFN-γ treatment detected by flow cytometry (F) and immunofluorescence (G). Scale bar, 10 μm. (H) Western blot analysis of CD47 levels in A549 cells transfected with IRF-1-specific or scramble shRNA. Left: Representative image; right: quantitative analysis. Data from 3 independent experiments were presented as means ± SDs. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Expressing, Sequencing, Binding Assay, Reporter Assay, Luciferase, Transfection, shRNA, Flow Cytometry, Immunofluorescence, Western Blot

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: IFN-γ promotes lung cancer cell metastasis by upregulating CD47 expression in vitro (A) Scratch-wound healing assay in A549 cells (A549-WT) and CD47-knockout A549 cells (A549-CD47-KO) were treated with/without IFN-γ. (B) Transwell assay in A549 cells (A549-WT) and A549-CD47-KO cells were treated with/without IFN-γ. Data from 3 independent experiments were presented as means ± SDs. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bar, 200 μm.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Expressing, In Vitro, Wound Healing Assay, Knock-Out, Transwell Assay

Journal: Molecular Therapy Oncolytics

Article Title: Human lung adenocarcinoma CD47 is upregulated by interferon-γ and promotes tumor metastasis

doi: 10.1016/j.omto.2022.04.011

Figure Lengend Snippet: IFN-γ promotes human lung cancer cell metastasis in immunodeficient mice by upregulating CD47 (A) A549 cells (A549-WT) and A549-CD47-KO were engrafted in the lungs of the BALB/c-nude mice. Three weeks post-engraftment, the mice were randomly divided into 2 groups. One group was administered with IFN-γ (10 ng/mouse, injected once every 2 days) (A549-WT + IFN-γ; A549-CD47-KO + IFN-γ), and the other group without IFN-γ injection was served as control (A549-WT; A549-CD47-KO). After 5 weeks, the mice were sacrificed to analyze tumor growth and metastasis. (B and C) H&E staining and human CD47 immune staining in mouse lungs (B) and livers (C). In both (B) and (C), left: representative images; right: quantification of tumor nodules. Data were presented as means ± SDs. ∗p < 0.05, ∗∗∗p < 0.001. NS, no significance.

Article Snippet: Primary antibodies against CD47 (AF4670, R&D Systems, Minneapolis, MN, USA), PD-L1 (#13684, Cell Signaling Technology) and IRF1 (11335-1-AP, Proteintech) were used.

Techniques: Injection, Control, Staining

Journal: Oncology Letters

Article Title: CD47 as a potential prognostic marker for oral leukoplakia and oral squamous cell carcinoma

doi: 10.3892/ol.2018.8520

Figure Lengend Snippet: Expression of CD47 in oral squamous cell carcinoma cell lines and normal oral keratinocytes, as assessed by immunofluorescence. CD47 was expressed in (A1) Tca8113, (B1) Cal-27 and (C1) SCC-9 cells. (D1) Weak positive staining was observed in normal oral keratinocytes. No expression of CD47 was observed in the control cells (A2-D2; magnification, ×400). CD47, cluster of differentiation 47.

Article Snippet: The sections were then incubated at 4°C overnight in a diluted solution (dilution, 1:200) of monoclonal mouse anti-human CD47 (cat. no. MAB4670; R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: Oncology Letters

Article Title: CD47 as a potential prognostic marker for oral leukoplakia and oral squamous cell carcinoma

doi: 10.3892/ol.2018.8520

Figure Lengend Snippet: Effect of CD47 antibody on the proliferation of Cal-27 cells. The proliferation of Cal-27 cells was inhibited by 5 and 10 µg/ml CD47 antibody. *P<0.05, **P<0.01 and ***P<0.001 vs. control group. CD47, cluster of differentiation 47; OD, absorbance; CCK-8, Cell Counting kit-8.

Article Snippet: The sections were then incubated at 4°C overnight in a diluted solution (dilution, 1:200) of monoclonal mouse anti-human CD47 (cat. no. MAB4670; R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Control, CCK-8 Assay, Cell Counting

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Molecular simulation of ligand, TSP1, interaction with the receptor (CD47) in a solvent-free environment. Slightly open conformation of the 4N1K hydrophobic cleft (RFYVVMWK peptide region, pink) on TSP1 C-terminal domain was modeled via Molecular Operating Environment (MOE) to participate in interaction with the N-terminus of the model of the soluble extracellular part of the CD47 receptor (shown in two mode views, ( A , B ), as identified experimentally, in addition to the two putative TSP1:CD47 interaction regions (1) and (2), respectively, as recognized earlier. Light green: residues of TSP1 involved in binding.

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Solvent, Binding Assay

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Qualitative and quantitative confirmation of association between TSP1 and the recombinant human CD47 peptide (rh-CD47p). ( A ) Representative image of immunoblots of combination of monomer TSP1 (mTSP1) and rh-CD47p incubated at 37 °C for 60 min in a cell-free setting. Lane 1: molecular weight marker; 2: mTSP1 control; 3: rh-CD47p control; 4: mTSP1+rh-CD47p mixture; 5: TSP1 blocking antibody alone (TSP1 mAb 301221); 6. rh-CD47p+TSP1 blocking antibody; 7: mTSP1+TSP1 blocking antibody; and 8: mTSP1+rh-CD47p mixture with TSP1 blocking antibody. mTSP1 control (Lane 2) is present as a clear band in red fluorescence at 180 KDa; rh-CD47p control (Lane 3) is presented as a smear and a widely spread band in green fluorescence at approximately 60 KDa due to glycosylation; the mixture of mTSP1 and rh-CD47p without TSP1 blocking antibody at 240 KDa (Lane 4) or stacked up greatly higher than 300 KDa, following treatment with TSP1 blocking antibody (Lane 8), are presented in red fluorescence. Quantitative determination of binding of rh-CD47p to mTSP1 was performed via modified ELISA. A fixed amount of rh-CD47p was co-incubated with mTSP1 or trimeric TSP1 (tTSP1) at 37 °C for 60 min in a cell-free setting. Unbound rh-CD47p was collected and subject to human CD47 ELISA. Free rh-CD47p, normalized to control, increased as the amount of mTSP1 ( B ) or tTSP1 ( D ) decreased in the mixtures (one-way ANOVA followed by a Bonferroni posthoc test; Panel B: n = 7, *** p < 0.0001, mTSP1:rh-CD47p 1:1 to 1:6 vs. rh-CD47p control, mTSP1:rh-CD47p 1:1 vs. 1:3 and 1:6, and mTSP1:rh-CD47p 1:3 vs. 1:6; Panel D, n = 5, *** p < 0.001, tTSP1:rh-CD47p 1:3 to 1:9 vs. rh-CD47p control, tTSP1:rh-CD47p 1:3 vs. 1:9; n.s., no significance, tTSP1:rh-CD47p 1:3 vs. 1:6 and 1:6 vs. 1:9). Correlation between DRP doses, expressed in a molar ratio of TSP1 to rh-CD47p, and the bound rate of DRP was constructed for combination with mTSP1 ( C ) and tTSP1 ( E ), respectively (Panel C, p < 0.0001; Panel E, p < 0.01).

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Western Blot, Incubation, Molecular Weight, Marker, Control, Blocking Assay, Fluorescence, Glycoproteomics, Binding Assay, Modification, Enzyme-linked Immunosorbent Assay, Construct

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Potency of acetylcholine, measured in EC 50 (mean ± SEM, nM), in recombinant human CD47 peptide (rh-CD47p) pretreatment and post-treatment cohorts.

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Control

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Pretreatment with recombinant human CD47 peptide (rh-CD47p) effectively prevents TSP1-blunted vasodilation. rh-CD47p was administered 15 min prior to incubation of TSP1. ( A ) Concentration-response curves of isolated mouse thoracic aorta exposed to monomeric TSP1 (mTSP1) alone or combinations of mTSP1 and rh-CD47p in two molar ratios. mTSP1-induced impairment of vasodilation was prevented via pretreatment with rh-CD47p (two-way ANOVA followed by a Bonferroni posthoc test; n = 3–6, *** p < 0.0001, mTSP1 vs. control; * p < 0.05, mTSP1:rh-CD47p 1:2 vs. control; ** p < 0.01, mTSP1:rh-CD47p 1:1 vs. control; †† p < 0.01 and ††† p < 0.001, mTSP1:rh-CD47p 1:1 or 1:2 vs. mTSP1; ‡‡ p < 0.01, and ‡‡‡ p < 0.001, mTSP1:rh-CD47p 1:1 vs. 1:2). ( B ) Percentages of relaxation for all treatment groups at 10 μM acetylcholine are presented as box and whisker plots showing median, 1st quartile, 3rd quartile, and 95% CI. Significant difference in relaxation was found between groups treated with mTSP1 alone and the combination of mTSP1 and rh-CD47p ( n = 3–6, *** p < 0.001, mTSP1 vs. control; †† p < 0.01, mTSP1:rh-CD47p 1:1 vs. mTSP1; ††† p < 0.001, mTSP:rh-CD47p 1:2 vs. mTSP1). ( C ) Concentration-response curves of isolated mouse thoracic aorta exposed to trimeric TSP1 (tTSP1) alone or the combination of tTSP1 and rh-CD47p in a single molar ratio. tTSP1-induced impairment of vasodilation was completely blocked via pretreatment with rh-CD47p (two-way ANOVA followed by a Bonferroni posthoc test; n = 4, *** p < 0.0001, TSP1 vs. control; ### p < 0.0001, TSP1:rh-CD47p 1:3 vs. TSP1; p > 0.05, TSP1:rh-CD47p 1:3 vs. control). ( D ) Percentages of relaxation for all treatment groups at 10 μM acetylcholine are presented as box and whisker plots showing median, 1st quartile, 3rd quartile, and 95% CI. rh-CD47p treatment protected the vessel from substantial inhibition of vasodilation by native TSP1 ( n = 4, *** p < 0.0001, tTSP1 vs. control; ### p < 0.0001, tTSP1:rh-CD47p 1:3 vs. tTSP1).

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Incubation, Concentration Assay, Isolation, Control, Whisker Assay, Inhibition

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Post-treatment with recombinant human CD47 peptide (rh-CD47p) exerts restorative effect on injury to vasodilation by monomeric TSP1 (mTSP1). mTSP1 was incubated 15 min prior to coincubation with rh-CD47p for 45 min. ( A ) Concentration-response curves of isolated mouse thoracic aorta exposed to mTSP1 alone or the combination of mTSP1 and rh-CD47p in two different molar ratios. Substantial injury to vasodilation caused by mTSP1 was restored to control level (two-way ANOVA followed by a Bonferroni posthoc test; n = 4, *** p < 0.001, mTSP1 vs. control; ** p < 0.01, mTSP1:rh-CD47p 1:2 vs. control; † p < 0.05, †† p < 0.01, and ††† p < 0.0001, mTSP1:rh-CD47p 1:1 or 1:2 vs. mTSP1; ‡ p < 0.05, mTSP1:rh-CD47p 1:1 vs. 1:2). ( B ) Percentages of relaxation at 10 μM acetylcholine are presented as box and whisker plots showing median, 1st quartile, 3rd quartile, and 95% CI. Two doses of rh-CD47p brought impaired vasorelaxation close to control ( n = 4, *** p < 0.0001, TSP1 vs. control; † p < 0.05, mTSP1:rh-CD47p 1:1 vs. mTSP1; †† p < 0.01, mTSP1:rh-CD47p 1:2 vs. mTSP1).

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Incubation, Concentration Assay, Isolation, Control, Whisker Assay

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Post-treatment with recombinant human CD47 peptide (rh-CD47p) in low and high molar ratios differentially restored trimeric TSP1 (tTSP1)-blunted vasodilation. Acute exposure to tTSP1 for an initial 15 min was followed by 45 min coincubation with rh-CD47p. ( A ) Concentration-response curves of isolated mouse thoracic aorta exposed to tTSP1 alone or the combination of tTSP1 and rh-CD47p in low molar ratios (tTSP1:rh-CD47p 1:3 and 1:4). Low doses of rh-CD47p post-treatment ameliorated tTSP1-impaired vasodilation with limited efficacy (two-way ANOVA followed by a Bonferroni posthoc test; n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001, tTSP1 alone, tTSP1:rh-CD47p 1:3 or 1:4 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001, tTSP1:rh-CD47p 1:3 or 1:4 vs. tTSP1). ( B ) Concentration-response curves of isolated mouse thoracic aorta exposed to tTSP1 alone or the combination of tTSP1 and rh-CD47p in high molar ratios (tTSP1:rh-CD47p 1:5 and 1:6). A total reversion of tTSP1-injured vasodilation was consistently observed in rh-CD47p post-treatment in high molar ratios (two-way ANOVA followed by a Bonferroni posthoc test; n = 4, *** p < 0.001, tTSP1 vs. control; # p < 0.05, ## p < 0.01, and ### p < 0.001, tTSP1:rh-CD47p 1:5 or 1:6 vs. tTSP1). ( C ) Percentages of maximal relaxation in response to 10 μM acetylcholine are presented as box and whisker plots showing median, 1st quartile, 3rd quartile, and 95% CI. Significantly reduced vasorelaxation observed in tTSP1-treated alone no longer existed after receiving low or high doses of rh-CD47p treatment ( n = 3–4, *** p < 0.001, tTSP1 vs. control; # p < 0.05, tTSP1:rh-CD47p 1:3 vs. tTSP1; ### p < 0.001, tTSP1:rh-CD47p 1:4 up to 1:6 vs. tTSP1).

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Concentration Assay, Isolation, Control, Whisker Assay

Journal: Biomedicines

Article Title: Novel Pharmaceutical Strategy for Selective Abrogation of TSP1-Induced Vascular Dysfunction by Decoy Recombinant CD47 Soluble Receptor in Prophylaxis and Treatment Models

doi: 10.3390/biomedicines9060642

Figure Lengend Snippet: Schematics illustrating the putative alleviation mechanism of TSP1-impaired vasodilation via administration of recombinant human CD47 peptide (rh-CD47p). Under pathological conditions, excessive TSP1 is present in circulation or acutely excreted upon damage to the vascular wall. Under either pathological condition, excessive TSP1 significantly alters vascular tone via binding to its cognate receptor CD47 expressed on vascular cells, presenting as inhibited endothelium-dependent vasodilation. rh-CD47p, containing the TSP1 binding domain, was given prior to or after exposure to TSP1 to interfere with the communication between TSP1 and endothelial CD47, showed in the panel of the rh-CD47p treatment model. Since the signaling cascade initiated upon the interaction between TSP1 and endothelial CD47, TSP1-impaired endothelium-dependent vasodilation was expected to be prevented or restored as rh-CD47p was administered prior to or after the TSP1 challenge, respectively.

Article Snippet: Finally, the levels of rh-CD47p in the collected filtrates were quantified via a DuoSet ® ELISA kit for human CD47 (R&D Systems, Minneapolis, MN, USA) per the manufacturer’s instructions ( n = 5–7).

Techniques: Recombinant, Binding Assay