sev  (ATCC)

Journal: bioRxiv

Article Title: A Stable Reporter Cell Line to Study Respiratory Syncytial Virus NS2-Mediated Inhibition of IFNB Promoter Activation

doi: 10.64898/2026.02.24.706909

Figure Lengend Snippet: In A) A549 cells were infected with RSV A2 (MOI=3; 8 hrs) before analysis of WCE by immunoblotting using an anti-RSV NS1/NS2 antibody (left panel). The specificity of the anti-RSV NS1/NS2 antibody was confirmed by transfection of siRNA-targeting NS1 or NS2 before infection with RSV A2 (right panel). Anti-actin was used as a loading control. B) A549 cells were transiently transfected with an empty vector or the hNS2-pcDNA5 plasmid containing a humanized version of the RSV Long NS2 coding sequence and analyzed by immunoblotting using anti-RSV NS1/NS2 antibodies. C) A549 cells were co-transfected with an IFNBprom-pGL3 firefly luciferase plasmid, a pRL-null renilla luciferase control plasmid (pRL-Null, Promega) and increasing amounts of hNS2-pcDNA5 plasmid before infection with Sendai virus (SeV) at 80HAU/10 6 cells. Luciferase activities were expressed as fold activation over the corresponding NS condition after normalization with renilla luciferase activities. Data (mean ±SEM; n=3) were analyzed using a two-tailed paired Student’s t-test.

Article Snippet: Alternatively, subconfluent monolayers of cells were infected with Sendai virus (SeV) Cantell strain (Charles River Laboratories) at 40 hemagglutinin units (HAU)/10 6 cells in the minimum volume of serum free medium.

Techniques: Infection, Western Blot, Transfection, Control, Plasmid Preparation, Sequencing, Luciferase, Virus, Activation Assay, Two Tailed Test

Journal: bioRxiv

Article Title: A Stable Reporter Cell Line to Study Respiratory Syncytial Virus NS2-Mediated Inhibition of IFNB Promoter Activation

doi: 10.64898/2026.02.24.706909

Figure Lengend Snippet: A) A549 cells transfected with either 3xFlag-pCMV-3Tag1a control or hNS2-3xFlag-pCMV-3Tag1a plasmids, followed by antibiotic selection to establish stable population and isolate control monoclonal cell lines (A549-ctrl) and NS2-expressing monoclonal cell lines (A549-Flag-NS2). The expression of Flag-tagged NS2 in the resulting monoclonal cells was confirmed by immunoblotting using anti-Flag antibody. Anti-actin was used as a loading control. Transiently transfected cells were used as positive control of Flag-NS2 expression. B) Pools of four A549-Ctrl or of A549-Flag-NS2 monoclonal cells were infected with Sendai virus (SeV) for the indicated times. IFNβ, TNF and SeV N transcript levels were quantified by RT-qPCR. C) A549-Ctrl and A549-Flag-NS2 cells were stimulated with recombinant human IFNβ for the indicated times. WCE were analyzed by immunoblotting using antibodies against total STAT2, phosphorylated STAT2 (Tyr689). Anti-actin was used as a loading control.

Article Snippet: Alternatively, subconfluent monolayers of cells were infected with Sendai virus (SeV) Cantell strain (Charles River Laboratories) at 40 hemagglutinin units (HAU)/10 6 cells in the minimum volume of serum free medium.

Techniques: Transfection, Control, Selection, Expressing, Western Blot, Positive Control, Infection, Virus, Quantitative RT-PCR, Recombinant