Journal: Viruses

Article Title: Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

doi: 10.3390/v12020212

Figure Lengend Snippet: Detection of WNV NY99 NS4B with the anti-JEV NS4B antibody. Vero cells were mock-infected or infected with ( A ) JEV (MOI of 2) and WNV NY99 (MOI of 1), ( B ) WNV NY99 (MOI of 1) and DENV-2 (MOI of 2) or ( C ) DENV serotypes 1, 2, 3, 4 (MOI of 2) and harvested 24 h after infection. Vero cell lysates (50 μg) were separated by 4–12% SDS-PAGE and western blot analyses was performed with antibodies against JEV NS4B, flavivirus Env, DENV-2 NS4B and calnexin, as indicated on the right side of each panel. Calnexin served as an internal loading control and the presence of the Env protein was used to confirm flavivirus infection. Molecular weights (kDa) are given on the left side of each panel. Abbreviations: DENV-1: Dengue 1; DENV-2: Dengue 2; DENV-3: Dengue 3; DENV-4: Dengue 4; Env: Envelope.

Article Snippet: The anti-flavivirus dsRNA mouse monoclonal antibody (J2 monoclonal antibody, Cat#10010200) was purchased from the English & Scientific Consulting in Hungary.

Techniques: Infection, SDS Page, Western Blot, Control

Journal: Viruses

Article Title: Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

doi: 10.3390/v12020212

Figure Lengend Snippet: Colocalization of WNV NY99 NS4B with Env protein, NS1 protein, and viral dsRNA in WNV-infected cells. Vero cells were infected with WNV NY99 at a MOI of 1 ( a – i , m – o ) or mock infected ( j – l ). At 24 h after infection, cells were fixed with 3.7% paraformaldehyde and processed for immunofluorescence (IF). Each panel represents the co-localization of cells (100% co-localization) in 10 to 15 fields in two independent infection experiments, with an average infection efficiency of 15%. Antibodies used for immunostaining are depicted on the top of the panels. Nuclear DNA was stained with 4,6-diamidino-2-phenylindole (DAPI). Merged pictures are shown on the right ( c , f , i , l , o ). Slides were analyzed by confocal laser scanning microscopy. Confocal microscopy images were of optical slice thickness ~1 μm. Scale bar, 10 μm. Abbreviations: WNV: West Nile virus NY99 strain (WNV NY99 ); JEV: Japanese encephalitis virus; NS1: Non-structural protein 1; NS4B: Non-structural protein 4B.

Article Snippet: The anti-flavivirus dsRNA mouse monoclonal antibody (J2 monoclonal antibody, Cat#10010200) was purchased from the English & Scientific Consulting in Hungary.

Techniques: Infection, Immunofluorescence, Immunostaining, Staining, Confocal Laser Scanning Microscopy, Confocal Microscopy, Virus

Journal: Viruses

Article Title: Selective Reactivity of Anti-Japanese Encephalitis Virus NS4B Antibody Towards Different Flaviviruses

doi: 10.3390/v12020212

Figure Lengend Snippet: Hydrophobicity and multiple sequence alignment analyses of NS4B of different flaviviruses. ( A ) Summary of transmembrane prediction results generated by the SOSUI program showing the number, positions, residues, length, and type of each of the transmembrane helices. ( B ) Alignment summary scores after multiple sequence alignment of NS4B proteins of WNV and four serotypes of dengue against JEV using ClustalW2 program. ( C ) Complete NS4B amino acid alignments of the WNV NY99 and JEV Nakayama strains. Highlighted residues in different colors depict the predicted location of the four transmembrane helices. The boxed region depicts the highly conserved region among flaviviruses. Virus abbreviations and (Genbank accession number): DENV-1: Dengue 1 (EU848545.1); DENV-2: Dengue 2 (EF457904.1); DENV-3: Dengue 3 (M93130.1); DENV-4: Dengue 4 (AY947539.1); WNV: West Nile virus NY99 strain (YP_001527886); JEV: Japanese encephalitis (NP_775673).

Article Snippet: The anti-flavivirus dsRNA mouse monoclonal antibody (J2 monoclonal antibody, Cat#10010200) was purchased from the English & Scientific Consulting in Hungary.

Techniques: Sequencing, Generated, Virus