Journal:

Article Title: Andes and Prospect Hill Hantaviruses Differ in Early Induction of Interferon although Both Can Downregulate Interferon Signaling ▿

doi: 10.1128/JVI.02402-06

Figure Lengend Snippet: A. IRF-3 dimerization in ANDV- or PHV-infected primary cells. HMVEC-L cells were infected with virus at an MOI of 1 or treated with inactivated ANDV or PHV. Cell extracts were analyzed on a nondenaturing native gel followed by Western blotting with anti-IRF-3 antibody. Sendai virus infection of HMVEC-L cells was used as a positive control. IRF-3 dimer formation was evident in PHV-infected cells at 12 h postinfection, whereas in ANDV-infected cells it was barely detected even by 24 h postinfection. No IRF-3 dimerization was detected with either inactivated PHV or ANDV. B. IRF-3 nuclear translocation induced by PHV infection. PHV- or ANDV-infected HMVEC-L cells were examined at 24 h postinfection by double immunofluorescent staining using anti-IRF-3 antibody (left panels) and cross-reactive anti-N monoclonal antibody GB04-BF07 (middle panels). Right panels show the merged images. No nuclear staining of IRF-3 was observed in uninfected or cells treated with inactivated PHV or ANDV (data not shown).

Article Snippet: Sendai virus strain Cantell was obtained from ATCC and propagated in 10-day-old embryonated chicken eggs.

Techniques: Infection, Virus, Western Blot, Positive Control, Translocation Assay, Staining

Journal:

Article Title: Andes and Prospect Hill Hantaviruses Differ in Early Induction of Interferon although Both Can Downregulate Interferon Signaling ▿

doi: 10.1128/JVI.02402-06

Figure Lengend Snippet: A. ANDV and PHV glycoproteins inhibit IFN-β-induced Stat-1 nuclear translocation. Vero-E6 cells were transfected with plasmids expressing ANDV GPC, PHV GPC, or Ebola virus VP35 (as a control). At 48 h posttransfection, cells were treated with 1,000 U of IFN-β for 20 min, fixed, and double stained using antibodies detecting Stat-1 (left panels), ANDV glycoproteins, PHV glycoproteins, and Ebola virus VP35 (middle panels). Right panels show the merged images. B. ANDV glycoprotein expression does not result in a general block of cellular protein nuclear translocation. Vero-E6 cells were transfected with plasmids expressing HA-tagged IRF-3 and ANDV GPC. At 24 h posttransfection cells were infected with Sendai virus (row 2) or left uninfected (row 1). The cells were fixed at 40 h posttransfection and double stained using anti-HA monoclonal antibody detecting expressed IRF-3 (left panels) or rabbit anti-ANDV polyclonal sera (middle panels). Right panels show the merged images.

Article Snippet: Sendai virus strain Cantell was obtained from ATCC and propagated in 10-day-old embryonated chicken eggs.

Techniques: Translocation Assay, Transfection, Expressing, Virus, Control, Staining, Blocking Assay, Infection

Journal: medRxiv

Article Title: Clinical validation of a novel metagenomic nanopore sequencing method for detecting viral respiratory pathogens

doi: 10.64898/2026.02.06.26345651

Figure Lengend Snippet: Red indicates passed and black failed. RNA from murine respirovirus, orthoreovirus and Zika virus were spiked as cDNA synthesis and SISPA controls, MS2 phage was spiked as an extraction, cDNA synthesis and SISPA control. Within the derivation set MS2 was not required to pass QC due to poor performance of the laboratory MS2 stock; in both validation and derivation only detection of 2/3 virus spikes was required to pass QC.

Article Snippet: Extracted RNA (8μl) was treated with DNAse, as recommended, then spiked with three reverse transcription/amplification controls (1μl each, containing approximately 10 4 genome copies each of Zika virus (ATCC-VR-1838DQ), murine respirovirus (ATCC-VR-907DQ) and orthoreovirus (ATCC-VR-824DQ) (ATCC Manassas, Virginia, USA). cDNA synthesis was primed randomly using the 3’ terminal N 9 segment of custom oligonucleotide primers (5’-GAT-GAT-AGT-AGG-GCT-TCG-TCA-CNN-NNN-NNN N-3’; Integrated DNA Technologies, Leuven, Belgium), final concentration 5μM.

Techniques: Virus, cDNA Synthesis, Extraction, Control, Biomarker Discovery

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