anti rig1 antibody  (Proteintech)

Journal: Archives of Virology

Article Title: An NLR family member X1 mutation ( p.Arg707Cys ) suppresses hepatitis B virus infection in hepatocytes and favors the interaction of retinoic acid-inducible gene 1 with mitochondrial antiviral signaling protein

doi: 10.1007/s00705-024-06133-0

Figure Lengend Snippet: Effect of the NLRX1 mutation on MAVS/RIG1 and proinflammatory cytokines. Huh7-NTCP cells are transfected with WT NLRX1 or MT NLRX1 vector for 48 h. ( A ) Coimmunoprecipitation assay showing the effect of WT NLRX1 or MT NLRX1 on MAVS/RIG1 interaction. ( B ) Luciferase expression from the IFN-α, IL-6, or IFN-β promoter in Huh7-NTCP cells. All experiments were repeated three times independently. *, P < 0.05 vs. NC + HBV; **, P < 0.01 vs. NC + HBV; +, P < 0.05 vs. WT NLRX1 + HBV

Article Snippet: The primary antibodies were as follows: HBcAg (Santa Cruz, #sc-23947; 1:1000); HBsAg (Santa Cruz, #sc-53299; 1:1000); NLRX1 (Proteintech, Wuhan, China, #17215-1-AP; 1:2000); RIG1 (CST, #3743T; 1:2000); MAVS (Abcam, ab31334; 1:2000); p65 (Abcam, ab7970; 1:500); p-p65 (CST, #3033S; 1:1000); IRF7 (CST, #13014; 1:1000); p-IRF7 (CST, #12390; 1:1000); IRF3 (CST, #4302; 1:1000); p-IRF3 (CST, #29047; 1:1000); and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Proteintech, 60004-1-Ig, 1:8000).

Techniques: Mutagenesis, Transfection, Plasmid Preparation, Co-Immunoprecipitation Assay, Luciferase, Expressing

Journal: iScience

Article Title: Alphaherpesvirus manipulates retinoic acid metabolism for optimal replication

doi: 10.1016/j.isci.2024.110144

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-RIG1 , Proteintech , Cat# 20566-1-AP RRID: AB_10700006.

Techniques: Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: Cells

Article Title: Significant Variations in Double-Stranded RNA Levels in Cultured Skin Cells

doi: 10.3390/cells13030226

Figure Lengend Snippet: Fibroblasts express minimal amounts of dsRNA. DsRNA was assessed using the J2 antibody with dot blots and immunocytochemistry. ( A ) Three technical replicates of RNA were spotted on nitrocellulose probed with J2 and visualized using an HRP-coupled secondary antibody and CSPD star (Roche) detection. ( B ) Quantification of the luminescence signal in ( A ), three technical and two biological replicates (ANOVA with Tukey’s post hoc test). ( C ) Pictures 1–7, fibroblast cells; picture 8 (bottom right), A375 cells. The cells were fixed and stained with combinations of antibodies against dsRNA (J2, red) and dsRNA binding proteins (ADAR, RIG1, MDA5, PKR, and phosphoPKR) as well as TOMM20 (mitochondria) and DAPI (nucleus). In A375 control cells, the J2 signal is clearly visible throughout the cytoplasm, whereas it is undetectable in fibroblast.

Article Snippet: Primary antibodies against PKR (Abcam, ab32052, Cambridge, UK), RIG1 (Abcam, ab180675), MDA5 (Abcam, ab126630), ADAR (Atlas, B115763, Stockholm, Sweden) were used at a dilution of 1:5000 and anti-actin (Rabbit, Sigma; A2066) at 1:10,000.

Techniques: Immunocytochemistry, Staining, Binding Assay

Journal: Cells

Article Title: Significant Variations in Double-Stranded RNA Levels in Cultured Skin Cells

doi: 10.3390/cells13030226

Figure Lengend Snippet: Double-stranded RNA sensors. ( A ) mRNA levels of dsRNA sensors in A375 and C8161 cells assessed using RT-qPCR. Actin was used as a reference. **** p < 0.001, ns, not significant, t -test. Of note, increased ΔCt values reflect lower mRNA expression levels. ( B ) Protein levels of dsRNA sensors assessed using Western blotting. ** p < 0.01, ns, not significant, t -test. Blots are provided in . ( C ) Schematic diagram of dsRNA triggered pathways. Left, PKR recognizes short perfect RNA hybrids, dimerizes, and undergoes autophosphorylation. Phospho PKR induces global translational arrest via phosphorylation of eIF2α and leads to NF-κB mediated interferon signaling. Middle, the two Rig-like receptors (RLR) RIG1and MDA5 bind dsRNA with different specificity; RIG1 binds 5′ phosphate groups and polymerizes along dsRNA, whereas MDA5 binds long (500–1000bp) RNA hybrids. Both activate MAVS and lead to NF-κB mediated immune signaling and apoptosis. Right, dsRNA from nuclear DNA becomes edited by ADAR1 and is prevented from exiting the nucleus. Should the barrier be breached, MDA5 and PKR may become activated.

Article Snippet: Primary antibodies against PKR (Abcam, ab32052, Cambridge, UK), RIG1 (Abcam, ab180675), MDA5 (Abcam, ab126630), ADAR (Atlas, B115763, Stockholm, Sweden) were used at a dilution of 1:5000 and anti-actin (Rabbit, Sigma; A2066) at 1:10,000.

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Cells

Article Title: Significant Variations in Double-Stranded RNA Levels in Cultured Skin Cells

doi: 10.3390/cells13030226

Figure Lengend Snippet: Immunohistochemical staining of dsRNA and dsRNA sensor proteins. The two cell lines, A375 and C8161, were assessed with the indicated antibodies against MDA5, RIG1, ADAR, PKR, and dsRNA (J2). The left panel shows nuclear DAPI staining (blue), followed by the specific dsRNA sensor proteins (green) and dsRNA (J2) in red. The panel on the right displays the composite. Scale bars are 10 µm (left panel). In the right panel, the spatial overlap of signals is compared between A375 and C8161 cells (R 2 ). * p < 0.05, ** p < 0.01, **** p < 0.001, ns, not significant, t -test.

Article Snippet: Primary antibodies against PKR (Abcam, ab32052, Cambridge, UK), RIG1 (Abcam, ab180675), MDA5 (Abcam, ab126630), ADAR (Atlas, B115763, Stockholm, Sweden) were used at a dilution of 1:5000 and anti-actin (Rabbit, Sigma; A2066) at 1:10,000.

Techniques: Immunohistochemistry, Staining

Journal: Cells

Article Title: Significant Variations in Double-Stranded RNA Levels in Cultured Skin Cells

doi: 10.3390/cells13030226

Figure Lengend Snippet: Expression differences in components of dsRNA signaling pathways between cell lines. ( A ) Expression differences as established using DESeq of selected dsRNA sensors, including MDA5, PKR, ADAR, RIG1, OAS, RNaseL, and MAVS. * adjusted p -value < 0.05; *** adjusted p -value < 0.005. ( B ) Components enriched in the KEGG pathway ‘Rig-like receptor signaling’ in C8161 cells versus fibroblasts and A375 versus fibroblasts. +P, phosphorylation; +U, ubiquitination; −U, deubiquitination. Adjusted p -value < 0.05.

Article Snippet: Primary antibodies against PKR (Abcam, ab32052, Cambridge, UK), RIG1 (Abcam, ab180675), MDA5 (Abcam, ab126630), ADAR (Atlas, B115763, Stockholm, Sweden) were used at a dilution of 1:5000 and anti-actin (Rabbit, Sigma; A2066) at 1:10,000.

Techniques: Expressing