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Novus Biologicals sars cov 2 envelope 2 e protein

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 <t>μg/mL),</t> <t>2-E</t> (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.

Sars Cov 2 Envelope 2 E Protein, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+sars+cov+2+envelope+protein/Recombinant+SARS-CoV-2+Envelope+(Avi+Epitope+Tag)+His+(N-Term)+Avi-tag+Protein/pmc13083715-55-0-8
Average 94 stars, based on 6 article reviews

sars cov 2 envelope 2 e protein - by Bioz Stars, 2026-10

94/100 stars

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1) Product Images from "Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo"

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

Journal: Journal of Traditional and Complementary Medicine

doi: 10.1016/j.jtcme.2025.12.003

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.

Figure Legend 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.

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

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.

Figure Legend 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.

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

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.

Figure Legend 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.

Techniques Used: Western Blot, Control

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.

Figure Legend 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.

Techniques Used: Phospho-proteomics, Western Blot, Control

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.

Figure Legend 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.

Techniques Used: Phospho-proteomics, Western Blot, Control

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.

Figure Legend 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.

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

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

Techniques Used:

other:

Article Title: SARS-CoV-2 envelope protein regulates innate immune tolerance
Article Snippet: Cryopreserved vials of PBMCs, unique donors, were acquired (StemCell Technologies, Vancouver, BC, Canada), thawed according to manufacturer guidelines and resuspended in phosphate buffered saline (PBS) with 0.1% bovine serum albumin (BSA) and counted on the Countess III (Thermo Fisher Scientific, Waltham, MA, USA).

Recombinant:

Article Title: SARS-CoV-2 envelope protein regulates innate immune tolerance
Article Snippet: Forget-Me-Not EvaGreen qPCR Master Mix , Biotium , 31041. .. Recombinant SARS-CoV-2 Envelope protein , Novus Biologicals , NBP2-90986. .. Recombinant SARS-CoV-2 Envelope Protein , AbClonal , RP01263LQ.

Image Search Results

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:

The SARS-CoV-2 E protein is intracellularly located and colocalizes with the endoplasmic reticulum in hippocampal neurons. E protein location was tested in rat hippocampal cultures treated with the vehicle (Control) or the E protein (E protein, 0.6 µg/mL) for 24 h. Immunochemistry experiments were independently performed with two different antibodies, against the E protein and HisTag. Incubation with ER-Tracker allowed one to observe endoplasmic reticulum. ( A ) Representative bright field and immunofluorescence images of neurons treated with the E protein and nuclei (FITC|Hoechst) taken from young (6–8 DIV) and aged (20–22 DIV) hippocampal neurons using antibodies against the E protein (α-E protein Ab). Alongside is the quantitative age-independent analysis of immunofluorescence intensity levels (Optical density in arbitrary units) for control and E protein (E prot), using E protein antibody. The bars represent the mean ± SEM from 133 and 165 cells from five independent experiments. The scale bar is 10 µm. * p < 0.05 vs. control. ( B ) The same procedure using HisTag antibody (α-HisTag Ab). Data were taken from 190 and 207 cells from five independent experiments. The scale bar is 10 µm. * p < 0.05 vs. control. ( C ) Representative confocal fluorescence images from endoplasmic reticulum (ER-Tracker), nuclei (Hoechst), the E protein using E protein antibody (E prot FITC) and the merge of a 21 DIV neuron. The bars represent the Manders coefficient M1 (ER-Tracker overlapping FITC) and M2 (FITC overlapping ER-Tracker) mean ± SEM from 16 cells in three independent experiments. The scale bar is 5 µm.

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: The SARS-CoV-2 E protein is intracellularly located and colocalizes with the endoplasmic reticulum in hippocampal neurons. E protein location was tested in rat hippocampal cultures treated with the vehicle (Control) or the E protein (E protein, 0.6 µg/mL) for 24 h. Immunochemistry experiments were independently performed with two different antibodies, against the E protein and HisTag. Incubation with ER-Tracker allowed one to observe endoplasmic reticulum. ( A ) Representative bright field and immunofluorescence images of neurons treated with the E protein and nuclei (FITC|Hoechst) taken from young (6–8 DIV) and aged (20–22 DIV) hippocampal neurons using antibodies against the E protein (α-E protein Ab). Alongside is the quantitative age-independent analysis of immunofluorescence intensity levels (Optical density in arbitrary units) for control and E protein (E prot), using E protein antibody. The bars represent the mean ± SEM from 133 and 165 cells from five independent experiments. The scale bar is 10 µm. * p < 0.05 vs. control. ( B ) The same procedure using HisTag antibody (α-HisTag Ab). Data were taken from 190 and 207 cells from five independent experiments. The scale bar is 10 µm. * p < 0.05 vs. control. ( C ) Representative confocal fluorescence images from endoplasmic reticulum (ER-Tracker), nuclei (Hoechst), the E protein using E protein antibody (E prot FITC) and the merge of a 21 DIV neuron. The bars represent the Manders coefficient M1 (ER-Tracker overlapping FITC) and M2 (FITC overlapping ER-Tracker) mean ± SEM from 16 cells in three independent experiments. The scale bar is 5 µm.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Control, Incubation, Immunofluorescence, Fluorescence

The SARS-CoV-2 E protein induces cell death in aged cultures of rat hippocampal neurons but not in young cultures. Primary hippocampal neuron cultures for 6–8 DIV (young) and 20–22 DIV (aged) were treated for 24 h in the absence or presence of the SARS-CoV-2 E protein. Apoptosis was assessed 24 h later by means of staining with Annexin V. ( A ) Representative bright field and Annexin V inmunofluorescence images of young (upper) and aged (bottom) hippocampal cultures in the presence (E prot) or absence (Control) of the E protein (0.6 μg/mL). A bar represents 10 μm and applies to all photographs. ( B ) Percentage of neuron dead cells in control and E protein-treated cells in young and aged cultures. Values represent the mean ± SEM from 865, 880, 561 and 639 cells from seven independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. young cultures.

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: The SARS-CoV-2 E protein induces cell death in aged cultures of rat hippocampal neurons but not in young cultures. Primary hippocampal neuron cultures for 6–8 DIV (young) and 20–22 DIV (aged) were treated for 24 h in the absence or presence of the SARS-CoV-2 E protein. Apoptosis was assessed 24 h later by means of staining with Annexin V. ( A ) Representative bright field and Annexin V inmunofluorescence images of young (upper) and aged (bottom) hippocampal cultures in the presence (E prot) or absence (Control) of the E protein (0.6 μg/mL). A bar represents 10 μm and applies to all photographs. ( B ) Percentage of neuron dead cells in control and E protein-treated cells in young and aged cultures. Values represent the mean ± SEM from 865, 880, 561 and 639 cells from seven independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. young cultures.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Staining, Control

The SARS-CoV-2 E protein induces diverse cytosolic calcium responses in young and aged hippocampal neurons. Hippocampal neurons were loaded with fura2/AM and subjected to fluorescence imaging to monitoring changes in cytosolic calcium levels in response to E protein (2 μg/mL) acute treatment. Traces are representative recordings of fluorescence ratios in individual neurons for each type of response, ( A ) vehicle treatment (Control), ( B ) E protein treatment with lack of response (No response), ( C ) low calcium response (Low response), and ( D ) high calcium response (High response) in young (6–8 DIV) and aged (20–22 DIV) cultures. Neurons were identified by their morphology and N-Methyl-D-Aspartate response (NMDA, 100 µM). The pictures show bright field images (BF) and pseudocolor images of fluorescence ratios taken before (Basal) and after vehicle (Vehicle) or E protein (E prot) acute treatment in young (6–8 DIV) and aged (20–22 DIV) cultured neurons. The pseudocolor scale is shown at right. A bar represents 10 μm and applies to all photographs. ( E ) The bars represent the percentage of each type of response observed, according to the age of culture. ( F ) The bar plot represents the mean ± SEM of the “Fraction × ΔRatio” parameter, which is the product of the proportion of responsive cells and the maximum height ratio F340/F380 (Δ ratio) n = 33, 36, 58 and 96 cells from 4–12 independent experiments * p < 0.05 vs. control.

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: The SARS-CoV-2 E protein induces diverse cytosolic calcium responses in young and aged hippocampal neurons. Hippocampal neurons were loaded with fura2/AM and subjected to fluorescence imaging to monitoring changes in cytosolic calcium levels in response to E protein (2 μg/mL) acute treatment. Traces are representative recordings of fluorescence ratios in individual neurons for each type of response, ( A ) vehicle treatment (Control), ( B ) E protein treatment with lack of response (No response), ( C ) low calcium response (Low response), and ( D ) high calcium response (High response) in young (6–8 DIV) and aged (20–22 DIV) cultures. Neurons were identified by their morphology and N-Methyl-D-Aspartate response (NMDA, 100 µM). The pictures show bright field images (BF) and pseudocolor images of fluorescence ratios taken before (Basal) and after vehicle (Vehicle) or E protein (E prot) acute treatment in young (6–8 DIV) and aged (20–22 DIV) cultured neurons. The pseudocolor scale is shown at right. A bar represents 10 μm and applies to all photographs. ( E ) The bars represent the percentage of each type of response observed, according to the age of culture. ( F ) The bar plot represents the mean ± SEM of the “Fraction × ΔRatio” parameter, which is the product of the proportion of responsive cells and the maximum height ratio F340/F380 (Δ ratio) n = 33, 36, 58 and 96 cells from 4–12 independent experiments * p < 0.05 vs. control.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Fluorescence, Imaging, Control, Cell Culture

$The SARS-CoV-2 E protein induces Ca 2+ release from intracellular stores in young and aged hippocampal neurons. The release of Ca 2+ store content was tested in hippocampal neurons from short-term (6–8 DIV) and long-term (20–22 DIV) cultures. Cells were loaded with fura2/AM (4 µM) and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium. ( A ) The pictures show bright field (BF) and pseudocolor images of fluorescence ratios in basal conditions (Basal) and after vehicle (Vehicle) or E protein (E protein) treatment (2 μg/mL), in short-term (6–8 DIV) and long-term (20–22 DIV) cultured neurons. The bar represents 10 μm and applies to all photographs. Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( B ) The bars correspond to the average (mean ± SEM) fraction of cells responding to vehicle or E protein, ( C ) maximum height ratio F340/F380 (Δ ratio) in responsive cells, ( D ) and the product of both parameters, n = 42, 63, 60 and 59 cells from 17 independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. 6–8 DIV cultures. For ( B – D ) n = 42, 63, 60 and 59 cells from 17 independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. young (6–8 DIV) cultures.$

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: The SARS-CoV-2 E protein induces Ca 2+ release from intracellular stores in young and aged hippocampal neurons. The release of Ca 2+ store content was tested in hippocampal neurons from short-term (6–8 DIV) and long-term (20–22 DIV) cultures. Cells were loaded with fura2/AM (4 µM) and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium. ( A ) The pictures show bright field (BF) and pseudocolor images of fluorescence ratios in basal conditions (Basal) and after vehicle (Vehicle) or E protein (E protein) treatment (2 μg/mL), in short-term (6–8 DIV) and long-term (20–22 DIV) cultured neurons. The bar represents 10 μm and applies to all photographs. Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( B ) The bars correspond to the average (mean ± SEM) fraction of cells responding to vehicle or E protein, ( C ) maximum height ratio F340/F380 (Δ ratio) in responsive cells, ( D ) and the product of both parameters, n = 42, 63, 60 and 59 cells from 17 independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. 6–8 DIV cultures. For ( B – D ) n = 42, 63, 60 and 59 cells from 17 independent experiments. * p < 0.05 vs. control; # p < 0.05 vs. young (6–8 DIV) cultures.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Fluorescence, Imaging, Cell Culture, Control

$The SARS-CoV-2 E Protein prevents CPA liberation of calcium stores in aged hippocampal neurons. The release of Ca 2+ store content was tested with CPA (10 µM) in hippocampal neurons from short-term (Young, 6–8 DIV) and long-term (Aged, 20–22 DIV) cultures. Cells were loaded with fura2/AM and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium (Ca0). ( A ) The pictures show pseudocolor images of fluorescence ratios in basal conditions (Basal), after vehicle (Vehicle) or E protein (E Prot) acute treatment (2 μg/mL) and after CPA perfusion (CPA) in young and aged cultures. The bars represent 10 μm. Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( B ) The line series display fluorescence paired corresponding to the CPA response after vehicle (Vehicle) or E protein (E Prot) treatment in young and aged cultures. ( C ) The bars correspond to the average (mean ± SEM) product of the fraction of cells responding and the maximum height ratio F340/F380 (Δ ratio). For ( B , C ) n = 42, 25, 52 and 53 from 16 independent experiments. * p < 0.05 vs. control; *** p < 0.001; # p < 0.05 vs. 6–8 DIV cultures.$

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: The SARS-CoV-2 E Protein prevents CPA liberation of calcium stores in aged hippocampal neurons. The release of Ca 2+ store content was tested with CPA (10 µM) in hippocampal neurons from short-term (Young, 6–8 DIV) and long-term (Aged, 20–22 DIV) cultures. Cells were loaded with fura2/AM and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium (Ca0). ( A ) The pictures show pseudocolor images of fluorescence ratios in basal conditions (Basal), after vehicle (Vehicle) or E protein (E Prot) acute treatment (2 μg/mL) and after CPA perfusion (CPA) in young and aged cultures. The bars represent 10 μm. Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( B ) The line series display fluorescence paired corresponding to the CPA response after vehicle (Vehicle) or E protein (E Prot) treatment in young and aged cultures. ( C ) The bars correspond to the average (mean ± SEM) product of the fraction of cells responding and the maximum height ratio F340/F380 (Δ ratio). For ( B , C ) n = 42, 25, 52 and 53 from 16 independent experiments. * p < 0.05 vs. control; *** p < 0.001; # p < 0.05 vs. 6–8 DIV cultures.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Fluorescence, Imaging, Control

$Preceding calcium stores depletion with CPA to Thapsigargin prevents SARS-CoV-2 E protein effect regardless of age. Hippocampal neurons were loaded with fura2/AM and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium (Ca0). The release of Ca 2+ store content was tested with CPA (10 µM) and thapsigargin (Tg, 1 µM). Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( A ) Cells were exposed to acute treatment with the E protein (2 μg/mL) after CPA perfusion. The pictures show pseudocolor images of fluorescence ratios in basal conditions (Basal), CPA perfusion (CPA), and contiguous E protein treatment (E prot). The bars correspond to the E protein response as the product of the fraction of cells responding to treatment and the ratio of maximum height average (mean ± SEM) from cells before CPA perfusion (Pre CPA) or after (Post CPA). n = 12 and 17 cells from three independent experiments * p < 0.05 vs. control. ( B ) Cells were loaded with thapsigargin for 15 min and exposed to acute treatment with E protein (2 μg/mL). Pictures show bright field (BF) and pseudocolor images of fluorescence ratios in basal conditions (Basal) and after E protein treatment (E prot). The bars correspond to the E protein response as the product of the fraction of cells responding and the ratio of maximum height average (mean ± SEM) from cells previously loaded with Tg (Tg) or not (∅Tg). n = 64 and 90 from seven independent experiments. * p < 0.05 vs. control. The bar represents 10 μm and applies to all photographs.$

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: Preceding calcium stores depletion with CPA to Thapsigargin prevents SARS-CoV-2 E protein effect regardless of age. Hippocampal neurons were loaded with fura2/AM and subjected to fluorescence Ca 2+ imaging in external Ca 2+ -free medium (Ca0). The release of Ca 2+ store content was tested with CPA (10 µM) and thapsigargin (Tg, 1 µM). Traces are representative recordings of fluorescence ratios in individual neurons identified by their morphology. ( A ) Cells were exposed to acute treatment with the E protein (2 μg/mL) after CPA perfusion. The pictures show pseudocolor images of fluorescence ratios in basal conditions (Basal), CPA perfusion (CPA), and contiguous E protein treatment (E prot). The bars correspond to the E protein response as the product of the fraction of cells responding to treatment and the ratio of maximum height average (mean ± SEM) from cells before CPA perfusion (Pre CPA) or after (Post CPA). n = 12 and 17 cells from three independent experiments * p < 0.05 vs. control. ( B ) Cells were loaded with thapsigargin for 15 min and exposed to acute treatment with E protein (2 μg/mL). Pictures show bright field (BF) and pseudocolor images of fluorescence ratios in basal conditions (Basal) and after E protein treatment (E prot). The bars correspond to the E protein response as the product of the fraction of cells responding and the ratio of maximum height average (mean ± SEM) from cells previously loaded with Tg (Tg) or not (∅Tg). n = 64 and 90 from seven independent experiments. * p < 0.05 vs. control. The bar represents 10 μm and applies to all photographs.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Fluorescence, Imaging, Control

Dynamics of SARS-CoV-2 E protein translocation to endomembranes in hippocampal neurons. The dynamics of E protein location were tested in rat hippocampal cultures treated with the E protein (E protein, 2 µg/mL) for 5, 30 and 240 min (4 h) in ( A ) young (6–8 DIV) and ( B ) aged (20–22 DIV) rat hippocampal neurons. Then, cells were fixed and immunochemistry experiments were performed using E protein antibodies and confocal microscopy. Representative immunofluorescence images and the corresponding bright field images are shown. The data are representative of three similar experiments. The scale bar is 10 µm.

Journal: International Journal of Molecular Sciences

Article Title: SARS-CoV-2 Viroporin E Induces Ca 2+ Release and Neuron Cell Death in Primary Cultures of Rat Hippocampal Cells Aged In Vitro

doi: 10.3390/ijms25126304

Figure Lengend Snippet: Dynamics of SARS-CoV-2 E protein translocation to endomembranes in hippocampal neurons. The dynamics of E protein location were tested in rat hippocampal cultures treated with the E protein (E protein, 2 µg/mL) for 5, 30 and 240 min (4 h) in ( A ) young (6–8 DIV) and ( B ) aged (20–22 DIV) rat hippocampal neurons. Then, cells were fixed and immunochemistry experiments were performed using E protein antibodies and confocal microscopy. Representative immunofluorescence images and the corresponding bright field images are shown. The data are representative of three similar experiments. The scale bar is 10 µm.

Article Snippet: SARS-CoV-2 Envelope His-Avi Tag Recombinant Protein (RP-87682) and SARS Envelope Protein Polyclonal Antibody (PA1-41158) were obtained from Invitrogen (Waltham, MA, USA).

Techniques: Translocation Assay, Confocal Microscopy, Immunofluorescence

Journal: iScience

Article Title: SARS-CoV-2 envelope protein regulates innate immune tolerance

doi: 10.1016/j.isci.2024.109975

Figure Lengend Snippet:

Article Snippet: Cells were treated with either media or SARS-CoV-2 Envelope Protein (Novus Biologicals) at 1:2 titrated concentrations ranging from 4 μg/mL to 250 ng/mL for 16 h hTLR2, TLR2 agonist Heat killed-Listeria (InvivoGen), was used as a TLR2 positive control at a concentration of 10ˆ9 cells/mL was used. hTLR4, TLR4 agonist LPS (InvivoGen), was used as a TLR4 positive control at a concentration of 1 μg/mL.

Techniques: Recombinant, Selection, Isolation, Software

Review

sars cov 2 envelope 2 e protein (Novus Biologicals)

Structured Review

Images

1) Product Images from "Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo"

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