sars cov 2 envelope 2 e protein  (Novus Biologicals)

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits NO, IL-6, and TNF-α production in RAW264.7 and MH-S cells. The cells were treated with JGF (50, 100, 150, 300, 600 μg/mL), 2-E (0.1 μM), DXT (10 μM), or LPS (0.1 μg/mL) for 24 h. ( A ) Cell viability was evaluated using crystal violet. ( B ) NO production was measured using the Griess assay. ( C-D ) IL-6 ( C ) and TNF-α ( D ) levels were determined by ELISA. EC 50 was calculated by CompuSyn software. Data was presented as mean ± standard deviation (SD) for groups (n = 3). Significant differences are denoted as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Griess Assay, Enzyme-linked Immunosorbent Assay, Software, Standard Deviation

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: Components of JGF inhibit 2-E-induced inflammation. The RAW264.7 and MH-S cells were co-treated with JGF compounds and 2-E for 24 h. ( A ) The 3D-HPLC fingerprint of JGF. Compound structures were sourced from the PubChem database. The detection wavelength ranged from 200 to 400 nm, and the injection volume was 20 μL. ( B ) Cell viability was evaluated using crystal violet. ( C ) NO production was measured using the Griess assay. ( D-E ) IL-6 ( D ) and TNF-α ( E ) levels were determined by ELISA. Data are presented as mean ± SD (n = 3). Statistical significance was determined relative to the 2-E group. Significant differences are denoted as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Injection, Griess Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF downregulates 2-E-induced iNOS and COX-2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 24 h. ( A ) Protein levels of iNOS and COX-2 in macrophages were measured by Western blot. ( B-C ) Quantification of iNOS and COX-2 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. The non-detected data showed as – or ND. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of STAT3 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JAK2 and STAT3 were measured by Western blot. ( B-C ) Quantification of phosphorylated JAK2 and STAT3 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of ERK1/2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JNK1/2, ERK1/2, p38, and p65 were measured by Western blot. ( B-C ) Quantification of phosphorylated JNK1/2, ERK1/2, p38, and p65 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF reduces the 2-E-induced proinflammatory cytokines in vivo . ( A ) The experimental scheme for mouse exposure. ( B-F ) Levels of IL-6 ( B ), TNF-α ( C ), IFN-γ ( D ), IL-1β ( E ), and IL-12 ( F ) in lung tissue and serum were measured by ELISA. Data are presented as mean ± SD (n = 9 for serum, except DXT group n = 6; n = 6 for lung tissue, except DXT group n = 3) ( G ) Representative histological images of lung tissue stained with H&E and IHC images for IL-6, TNF-α, and IL-1β expression. ( H-J ) Quantification of IL-6 ( H ), TNF-α ( I ), and IL-1β ( J ) positive areas using ImageJ (n = 3). Significant differences between the control (CTL) group and other groups are denoted by ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Significant differences between the 2-E group and 2-E + JGF group are indicated by #p < 0.05, ##p < 0.01, ###p < 0.001.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: Schematics showing the anti-inflammatory mechanism of JGF in 2-E-induced mice macrophages.

Article Snippet: SARS-CoV-2-envelope (2-E) protein (Cat# NBP2–90986) was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques:

Journal: Microbiology Spectrum

Article Title: Construction and immunogenicity of SARS-CoV-2 virus-like particle expressed by recombinant baculovirus BacMam

doi: 10.1128/spectrum.00959-24

Figure Lengend Snippet: Construction and characterization of SARS-CoV-2 VLP-expressing BacMam viruses VLP-BM and VLP-G-BM. (a) Schematic diagram of the pVLP-BM and pVLP-G-BM donor plasmids. pCMV, Cytomegalovirus promoter; S, M, and E, SARS-CoV-2 Wuhan spike, membrane, and envelope, respectively; pA, poly A sequence; pH, polyhedrin promoter; and VSV-G, vesicular stomatitis virus glycoprotein. (b) Identification of SARS-CoV-2 S, M, and E proteins in BacMam-transduced HEK-293 cells. Non-transduced cells (lane 1), cells transduced with VLP-BM (lane 2), or VLP-G-BM (lane 3) viruses [multiplicity of infection (MOI) = 20] were harvested at 48 hpt for Western blotting with spike-, membrane-, envelope-, or β-actin-specific antibodies. (c) Morphology of SARS-CoV-2 VLP produced in HEK-293 cells. Cells were transduced with VLP-BM virus at an MOI of 20 for VLP purification by sucrose gradient centrifugation at 48 hpt and subjected to transmission electron microscopy.

Article Snippet: Rabbit anti-SARS-CoV-2 Spike Protein S1/S2 (ThermoFisher Scientific), rabbit anti-SARS-CoV Membrane (M) Protein (Rockland), rabbit anti-SARS-CoV-2 Envelope (E) Protein (ThermoFisher Scientific), mouse anti-His-tag (Qiagen), and mouse anti-β-actin (Cell Signaling Technology) antibodies were used as primary antibodies.

Techniques: Expressing, Membrane, Sequencing, Virus, Transduction, Infection, Western Blot, Produced, Purification, Gradient Centrifugation, Transmission Assay, Electron Microscopy

Journal: Microbiology Spectrum

Article Title: Construction and immunogenicity of SARS-CoV-2 virus-like particle expressed by recombinant baculovirus BacMam

doi: 10.1128/spectrum.00959-24

Figure Lengend Snippet: VLP BacMam induced spike-specific immune response in mice. (a) Immunization schedule. BALB/c mice ( n = 8) were intramuscularly injected with PBS or 2.25 × 10 8 ifu of VLP BacMam and boosted at 14 days post-vaccination (dpv). Serum samples were collected 14 days after each immunization. Mice were euthanized at 28 dpv to collect bronchoalveolar lavage (BAL) samples. (b, c, and d) SARS-CoV-2 S1-specific IgG titers in sera were determined by ELISA after primary (b) and boosting (c) immunization. SARS-CoV-2 S1-spcific IgG titers in BAL after two immunizations were also determined (d).

Article Snippet: Rabbit anti-SARS-CoV-2 Spike Protein S1/S2 (ThermoFisher Scientific), rabbit anti-SARS-CoV Membrane (M) Protein (Rockland), rabbit anti-SARS-CoV-2 Envelope (E) Protein (ThermoFisher Scientific), mouse anti-His-tag (Qiagen), and mouse anti-β-actin (Cell Signaling Technology) antibodies were used as primary antibodies.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: Microbiology Spectrum

Article Title: Construction and immunogenicity of SARS-CoV-2 virus-like particle expressed by recombinant baculovirus BacMam

doi: 10.1128/spectrum.00959-24

Figure Lengend Snippet: VLP BacMam induced neutralization activity against SARS-CoV-2 spike pseudotyped lentivirus. (a) Identification of ACE2 protein expressed in ACE2-expressing HEK-293 (HEK-293-ACE2) cells. HEK 293 cells were transfected with pACE2-V5-His tag plasmid and selected with Puromycin. Survival cells under selective pressure were sub-cultured and the polyhistine-tagged ACE2 protein expression was demonstrated by Western blotting with an anti-His-tag antibody. The levels of β-actin were demonstrated by an anti-β-actin antibody. Lanes: 1, HEK-293 cell lysate; 2, HEK-293-ACE2 cell lysate. ( b and c) Reporter expression by SARS-CoV-2 spike pseudotyped lentivirus. HEK-293-ACE2 cells were transduced with mock, or SARS-CoV-2 spike pseudotyped lentivirus virus expressing both GFP and luciferase reporters. Cells were harvested at 48 hpt to demonstrate GFP expression by confocal microscopy (b) and luciferase activity by luciferase assay (c). (d) Neutralizing activity of VLP BacMam-vaccinated mouse sera against SARS-CoV-2 spike pseudotyped lentivirus transduction. SARS-CoV-2 spike pseudotyped lentivirus was treated with PBS- or VLP BacMam-injected mouse sera at a dilution of 1:20 and used for transducing HEK-293-ACE2 cells. The cells were harvested at 48 hpt for luciferase assay. The luciferase activities in the PBS group were set to 100.

Article Snippet: Rabbit anti-SARS-CoV-2 Spike Protein S1/S2 (ThermoFisher Scientific), rabbit anti-SARS-CoV Membrane (M) Protein (Rockland), rabbit anti-SARS-CoV-2 Envelope (E) Protein (ThermoFisher Scientific), mouse anti-His-tag (Qiagen), and mouse anti-β-actin (Cell Signaling Technology) antibodies were used as primary antibodies.

Techniques: Neutralization, Activity Assay, Expressing, Transfection, Plasmid Preparation, Cell Culture, Western Blot, Transduction, Virus, Luciferase, Confocal Microscopy, Injection