yellow fever virus ns1 protein  (Biosynth Carbosynth)

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Cell viability evaluation by the MTT assay. Flavivirus sNS1 was added to THP-1 cells at a concentration of 5 μg/mL for 24 h. DMSO 20% was used as a positive control of cell toxicity. Data are reported as mean ± SD of three independent experiments. *** p < 0.001.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: MTT Assay, Concentration Assay, Positive Control

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Factor Xa generations on infected media added to normal plasma. (A and B) FXaG in medium from cells treated or untreated with NS1 proteins (5ug/mL for 24 h) and triggered by 5 pM TF. FXa generation activity in commercial standardized pooled normal plasma (minimum 20 donors) added with medium from untreated cells (NT, dashed gray line), and treated cells with sNS1 from WNV (blue line), YFV (light-blue line), JEV (red line), TBV (dashed light-blue line), D1 (dashed green line), D2 (green line), D3 (dashed pink line), D4 (pink line). The stimulation with LPS (dashed yellow line) was used as a positive control. NT (dashed gray line) (C) FXaG triggered by 5 pM TF, in pooled normal plasma added with purified sNS1 proteins from D1 (dashed green line), WNV (blue line), JEV (red line), and NT (dashed gray line). All curves and samples of FXaG are virtually indistinguishable, indicating no direct activity in FXa generation of sNS1 proteins. (D) Peak baseline % compared to NT. Data were reported as SD ± mean of three replicates * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: Infection, Clinical Proteomics, Activity Assay, Positive Control, Purification

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Evaluation of the TF expression. Flavivirus sNS1 was added to THP-1 cells at a concentration of 5 μg/mL for 24 h. The stimulation with lipopolysaccharides (LPS) was used as a positive control. NT: untreated samples. (A) Gene expression analysis was performed by RT-qPCR, and data are reported as fold change vs NT. (B) TF content in cell supernatants was evaluated by an ELISA assay. Data are reported as SD ± mean of three replicates. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: Expressing, Concentration Assay, Positive Control, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Evaluation of the TF amount in purified MV fractions. THP-1 cells were stimulated with ATP (1 mM), and flavivirus sNS1 was added at a concentration of 5 μg/mL for 24 h. The stimulation with lipopolysaccharides (LPS) was used as a positive control. NT: untreated samples. TF content in purified MV fraction was evaluated by ELISA assay. Data are reported as SD ± mean of three replicates. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: Purification, Concentration Assay, Positive Control, Enzyme-linked Immunosorbent Assay

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Evaluation of IL-6 release. IL-6 concentration was evaluated in THP-1 cells treated with flavivirus sNS1, which was added to THP-1 cells at a concentration of 5 μg/mL for 24 h. The stimulation with lipopolysaccharides (LPS) was used as a positive control. NT: untreated samples. (A) IL-6 levels in cell supernatants and (B) in the MVs purified fraction, after ATP stimulation. Data are reported as SD ± mean of three replicates. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: Concentration Assay, Positive Control, Purification

Journal: ACS Omega

Article Title: Flavivirus Nonstructural Protein 1‑Driven Coagulation via Tissue Factor-Bearing Microvesicles: A Pilot Study

doi: 10.1021/acsomega.5c09129

Figure Lengend Snippet: Proposed model of the sNS1-driven modulation of coagulation in THP-1 cells. Schematic representation of how neurotropic flavivirus sNS1 induces IL-6 and TF expression, leading to the release of TF-bearing MVs (TF + NVs) and increased procoagulant, whereas hemorrhagic sNS1 proteins show reduced activation, corresponding to an anticoagulant profile.

Article Snippet: sNS1 protein from eight different Flaviviruses (West Nile Virus – WNV, Dengue Virus 1-D1, Dengue Virus 2 – D2, Dengue Virus 3 – D3, Dengue Virus 4 – D4, Tick-Borne Encephalitis Virus – TBV, Yellow Fever Virus – YFV and Japanese Encephalitis Virus – JEV; FLAVX4-NS1, the Native Antigen, UK) were used at a final concentration of 5 μg/mL , , for 24 h. All recombinant sNS1 proteins were commercially obtained (The Native Antigen Company, UK), expressed in human HEK293 cells, with a declared purity of >99% verified by SDS-PAGE and HPLC.

Techniques: Coagulation, Expressing, Activation Assay