Journal: Viruses

Article Title: Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses

doi: 10.3390/v12040400

Figure Lengend Snippet: List of polyclonal antibodies generated in this study.

Article Snippet: The purified rabbit polyclonal antibodies against the TOSV nucleoprotein N (T4) and non-structural protein NSs (T5) were developed by GenScript (Piscataway, The Netherlands).

Techniques: Generated, Purification, Virus

Journal: Viruses

Article Title: Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses

doi: 10.3390/v12040400

Figure Lengend Snippet: Characterization of rescued TOSV particles after amplification in BHK-21 cells and four successive passages in Vero cells. ( a ) Proteins from viral particles purified through sucrose gradient were separated by nonreducing SDS-PAGE and stained with Coomassie blue. ( b , c ) Viral particles of TOSV wt strain H4906, rTOSV, and Germiston virus (GERV) were analyzed by SDS-PAGE and Western blotting under non-reducing conditions. ( b ) Viral proteins were visualized using guinea pig polyclonal antisera raised against TOSV (T1) or GERV (GR1); ( c ) same as ( b ), except using either the guinea pig polyclonal antibodies T2 and T3 against TOSV G N and G C , respectively, or the rabbit polyclonal antibody T4 against the TOSV nucleoprotein N.

Article Snippet: The purified rabbit polyclonal antibodies against the TOSV nucleoprotein N (T4) and non-structural protein NSs (T5) were developed by GenScript (Piscataway, The Netherlands).

Techniques: Amplification, Purification, SDS Page, Staining, Virus, Western Blot

Journal: Viruses

Article Title: Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses

doi: 10.3390/v12040400

Figure Lengend Snippet: Infection of A549 cells by rTOSV and TOSV. ( a ) A549 cells were infected with TOSV and rTOSV (MOI ∼2) for 24 h and then immunostained for the intracellular TOSV structural proteins G N , G C , and N, using an anti-TOSV primary mouse polyclonal antibody and an AF568-coupled anti-mouse secondary monoclonal antibody (red). Nuclei were stained with Hoechst (blue), and samples were imaged by wide-field microscopy; ( b ) Same as in (a), except that infected cells were analyzed by flow cytometry; ( c ) A549 cells were exposed to TOSV and rTOSV at the indicated MOIs for 24 h. Infection was analyzed by flow cytometry after immunostaining against the three structural proteins G N , G C , and N. Error bars indicate SD. Simple linear regression test returned no significant difference between TOSV and rTOSV series of values; ( d ) Infection with TOSV and rTOSV (MOI ∼2) was monitored by flow cytometry in A549 cells over 48 h. Error bars indicate SD. Two-Way ANOVA with Geisser-Greenhouse correction and Sidak’s multiple comparison tests returned no significant difference between the TOSV and rTOSV series of values.

Article Snippet: The purified rabbit polyclonal antibodies against the TOSV nucleoprotein N (T4) and non-structural protein NSs (T5) were developed by GenScript (Piscataway, The Netherlands).

Techniques: Infection, Staining, Microscopy, Flow Cytometry, Immunostaining, Comparison

Journal: Viruses

Article Title: Novel Toscana Virus Reverse Genetics System Establishes NSs as an Antagonist of Type I Interferon Responses

doi: 10.3390/v12040400

Figure Lengend Snippet: A549 cells were infected with the wt TOSV strain, rTOSV, and rTOSVɸNSs. Infected cells were lysed 24 h later, and then analyzed by SDS-PAGE and Western blotting under non-reducing conditions. The guinea pig antibodies T2 and T3 were used to detect the TOSV structural proteins G N and G C , respectively, while the rabbit polyclonal antibodies T4 and T5 allowed the detection of the TOSV nucleoprotein N and non-structural protein NSs.

Article Snippet: The purified rabbit polyclonal antibodies against the TOSV nucleoprotein N (T4) and non-structural protein NSs (T5) were developed by GenScript (Piscataway, The Netherlands).

Techniques: Infection, SDS Page, Western Blot