antibody against zika virus zikv envelope protein  (Sino Biological)

Journal: Frontiers in Microbiology

Article Title: Deficient of glycosylation site in the envelop protein attenuated Zika virus replication in mosquito cells

doi: 10.3389/fmicb.2025.1603083

Figure Lengend Snippet: T156I mutation delays the replication of ZIKV in mosquito cells. (A,B) The wild type (WT) and T156I mutant (T156I) virus were subjected to infect mosquito C6/36 cells. The whole cell lysates were collected after 12–72 hpi to determine the expression level of E protein using western blotting assay. The GAPDH were used as the house-keeping gene for normalization. The grayscale of the bands in western blotting assay were analyzed by ImageJ software. Asterisks indicate significant differences at 24 hpi between cells infected by WT (orange column) and cells infected by T156I (green column). (C) Mosquito cells were infected by WT or T156I virus for 12 hpi and 72 hpi. E protein (green) and cell nucleic (DAPI, blue) were immuno-stained to analyze the expression of E protein. (D,E) Chloroquine and MG132 were added to the cells followed by WT or T156I infection. The whole cell lysates were collected after 24 hpi to determine the expression level of E protein using western blotting assay. The GAPDH were used for normalization. (F,G) Mosquito cells were infected by WT or T156I. The cell lysates were used to extract the total RNA and subjected to detect the mRNA level of R protein. The supernatants were used to determine the virus titer by plaque-forming assay in VeroE6 cells. (H) The morphology of plaques was scanned. All the results represent the mean value ± standard error of the mean pooled from three independent experiments with duplicated samples. Asterisks indicate significant differences between cells infected by WT (orange column) and cells infected by T156I (green column). Statistical analysis was performed with unpaired Student’s t -test, * p < 0.01, ** p < 0.005, *** p < 0.001.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation with primary antibody against Zika virus (ZIKV) envelope protein (SinoBiological, 40543-R029) overnight at 4 °C.

Techniques: Mutagenesis, Virus, Expressing, Western Blot, Software, Infection, Staining

Journal: Frontiers in Microbiology

Article Title: Deficient of glycosylation site in the envelop protein attenuated Zika virus replication in mosquito cells

doi: 10.3389/fmicb.2025.1603083

Figure Lengend Snippet: T156I mutation potentially weaken the stability of E-dimer but not antibody receptor binding affinity (A) The molecular simulation of the T156I mutation on the structure of ZIKV E protein. The red, yellow, and blue colors represent the domain I, domain II and domain III of the E protein which mediate the formation of E-dimer. (B) The protein model was evaluated using Ramachandran plot analysis. The results indicate that 92.3% of the amino acids are located in the most favorable region, 7.1% in additional allowed regions, 0.3% in generously allowed regions, and 0.3% in disallowed regions-totaling 100%, which confirming the reliability of the E protein structural model. (C) The potential interaction patterns at the 156I residue with neighboring amino acids. Yellow stick represents the hydrogen bonds, pink stick showed the weak C–H bonds, and the gray stick represent the two sets of hydrophobic interactions with surrounding residues. (D) The HDCOK program was used to perform a global docking simulation of the binding interface and interaction mode between the 156I mutant E protein and the ZIKV neutralizing antibody EDE2. (E) Revealed that the mutant complex forms seven sets of hydrogen bonds (gray), four sets of weak C–H bonds (yellow), and two sets of hydrophobic interactions (pink). (F) The potential interactions of I156 with surrounding residues in 2D analysis.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation with primary antibody against Zika virus (ZIKV) envelope protein (SinoBiological, 40543-R029) overnight at 4 °C.

Techniques: Mutagenesis, Binding Assay, Residue

Journal: Frontiers in Pharmacology

Article Title: Antiviral activity of eicosapentaenoic acid against zika virus and other enveloped viruses

doi: 10.3389/fphar.2025.1564504

Figure Lengend Snippet: Inhibition of ZIKV activity by EPA in vitro . The mRNA levels of ZIKV E (A) and NS1 (B) was evaluated in ZIKV-infected Vero cells treated with EPA via qRT−PCR. (C) The protein expression of ZIKV E was evaluated in ZIKV-infected Vero cells treated with EPA via Western blotting. (D) ZIKV activity was evaluated in ZIKV-infected Vero cells treated with EPA via cell-based immunodetection assay. (E) ZIKV activity was evaluated in ZIKV-infected Vero cells treated with EPA via indirect immunofluorescence analysis. All experiments were repeated three times. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n. s = not significant.

Article Snippet: The cells were incubated with a primary antibody against ZIKV E or NS1 (1:1000) (BioFront Technologies, United States) at 37°C for 1 h, followed by incubation with a horseradish peroxidase (HRP)-labeled secondary antibody (1:2000) (Cell Signalling Technology, United States).

Techniques: Inhibition, Activity Assay, In Vitro, Infection, Quantitative RT-PCR, Expressing, Western Blot, Immunodetection, Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Antiviral activity of eicosapentaenoic acid against zika virus and other enveloped viruses

doi: 10.3389/fphar.2025.1564504

Figure Lengend Snippet: EPA rapidly inhibited ZIKV replication and targeted the virus. Determination of mRNA levels of ZIKV E (A) and NS1 (B) in cells infected with EPA–ZIKV mixture pre-incubated for 0, 5, 10, 30, and 60 min. Determination of the mRNA levels of ZIKV E (C) and NS1 (D) in cells, which were divided into four groups according to the treatment of EPA and ZIKV. Cell-treatment group: EPA was incubated with Vero cells for 1 h before ZIKV infection. Virus-treatment group: ZIKV was pre-incubated with EPA for 30 min and subsequently inoculated into cells. Co-treatment group: EPA and ZIKV were simultaneously inoculated into cells. Post-treatment group: Cells were incubated with ZIKV for 1 h and washed before being treated with EPA. All experiments were repeated three times. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n. s = not significant.

Article Snippet: The cells were incubated with a primary antibody against ZIKV E or NS1 (1:1000) (BioFront Technologies, United States) at 37°C for 1 h, followed by incubation with a horseradish peroxidase (HRP)-labeled secondary antibody (1:2000) (Cell Signalling Technology, United States).

Techniques: Virus, Infection, Incubation

Journal: Frontiers in Pharmacology

Article Title: Antiviral activity of eicosapentaenoic acid against zika virus and other enveloped viruses

doi: 10.3389/fphar.2025.1564504

Figure Lengend Snippet: EPA disrupted the early stages of the ZIKV replication cycle. (A, B) Results of time-of-addition assay. The mRNA levels of ZIKV E (A) and NS1 (B) were evaluated via qRT-PCR. EPA was added at 0, 5, 10, 20, and 40 min after ZIKV infection or at 0, 0.5, 1, 2, and 4 h after 1 h of ZIKV infection. The effects of EPA on the binding of ZIKV to host cells (C, D) , its adsorption on host cells (E–G) , and its entry into host cells (H) are shown. The effects of EPA on the mRNA (C–F) and protein (G, H) expression of ZIKV E are shown. All experiments were repeated three times. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n. s = not significant.

Article Snippet: The cells were incubated with a primary antibody against ZIKV E or NS1 (1:1000) (BioFront Technologies, United States) at 37°C for 1 h, followed by incubation with a horseradish peroxidase (HRP)-labeled secondary antibody (1:2000) (Cell Signalling Technology, United States).

Techniques: Quantitative RT-PCR, Infection, Binding Assay, Adsorption, Expressing