cantell strain sev  (Charles River Laboratories)

Journal: bioRxiv

Article Title: The human DEAD-box protein DDX3X regulates host and viral mRNA translation during Sendai Virus infection

doi: 10.64898/2026.03.08.707086

Figure Lengend Snippet: (A) Overview of the different SeV RNA species. (B) Mean count of forward vs. reverse reads in 16hr SeV infected DDX3X PAR-CLIP samples aligned to SeV Cantell strain genome. (C) Distribution of RNAseq reads and DDX3X-crosslinked PAR-CLIP reads aligning to regions of the SeV Cantell strain genome at 16hpi (single RNAseq replicate of n = 6 and PAR-CLIP replicate of n = 2 shown). Transcription start sites and start codons of SeV genes, as well as a region presumed to correspond to SeV dcRNA, are shown. PAR-CLIP T-to-C conversions are indicated as T-to-C mis-matches. Colored sites possess a large proportion of reads with nucleotide mismatches, with the ratio of A,T,C,G shown by the ratio of green, red, blue and orange, respectively. (D) Effects of DDX3X knockdown on FLUC/RLUC signal from cells transfected with pGL3-Prom Firefly luciferase 5’UTR reporter constructs and pGL3-Renilla luciferase. The FLUC signal was normalised to RLUC signal for each sample, and median values of triplicates were then normalised to a pGL3-Prom control with no 5’UTR insert. Reporters containing SeV 5’UTRs were transfected into non-silencing control (NSC) or short-hairpin DDX3X knockdown (shDDX3X) HEK293T cells treated with doxycycline for 72h to induce DDX3X knockdown. P values are calculated via Mann-Whitney U test paired by biological replicate. * = p < 0.05, NS = p > 0.05.

Article Snippet: For SeV infections, cells were inoculated at a concentration of 80 Hemagglutinating-units/mL of Cantell-strain SeV purchased from Charles River Laboratories.

Techniques: Infection, RNA sequencing, Knockdown, Transfection, Luciferase, Construct, Control, MANN-WHITNEY

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

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Regulation and Dynamics of Interferon-beta Expression Revealed with a Knockin Reporter Mouse

doi: 10.4049/jimmunol.2400227

Figure Lengend Snippet: (A) BMDMs from I-TOMCAT KI/+ Rosa26 FSF-eYFP/+ mice treated with 10μg/mL DMXAA for 6, 24, 48, and 72hrs. (B) BMDMs from I-TOMCAT KI/KI, I-TOMCAT KI/KI Sting −/− , and I-TOMCAT KI/KI Mavs −/− mice. Treated for 6hrs with 4μg/mL CT-DNA, 4μg/mL ISD-45, 50 HAU/mL SeV Cantell strain (Charles River Lot# 7Y200221), or lipofectamine alone. (C) RT-PCR quantification of Ifnb1 mRNA transcript of BMDM from I-TOMCAT KI/KI, I-TOMCAT KI/KI Sting −/− , and I-TOMCAT KI/KI Mavs −/− mice treated for 6hrs as in (B). Statistics calculated by ordinary one-way ANOVA with Tukey’s Test. *p<0.05 **p<0.01 ****p<0.0001.

Article Snippet: Treated for 6hrs with 4μg/mL CT-DNA, 4μg/mL ISD-45, 50 HAU/mL SeV Cantell strain (Charles River Lot# 7Y200221), or lipofectamine alone. (C) RT-PCR quantification of Ifnb1 mRNA transcript of BMDM from I-TOMCAT KI/KI, I-TOMCAT KI/KI Sting −/− , and I-TOMCAT KI/KI Mavs −/− mice treated for 6hrs as in (B).

Techniques: Activation Assay, Expressing, Reverse Transcription Polymerase Chain Reaction