flavivirus ns1 protein pack  (Native Antigen Inc)

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: 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