antibodies rabbit polyclonal anti adar1  (Cell Signaling Technology Inc)

Journal: Oxidative Medicine and Cellular Longevity

Article Title: circHtra1/miR-3960/GRB10 Axis Promotes Neuronal Loss and Immune Deficiency in Traumatic Brain Injury

doi: 10.1155/2022/3522492

Figure Lengend Snippet: IGF-1 reduces circHtra1 via ADAR1. (a) IGF-1 is immunoprecipitated with htra1 in HEK cells, and this effect is blocked by AG1024. (b) circHtra1 expression in neurons treated with KA, IGF-1, ADAR1, and their inhibitors ( t = 4.919, 3.162, and 8.503, respectively, df = 4). (c, d) Expression of ADAR1 in neurons treated with KA, IGF-1, ADAR1, and their inhibitors ( t = 3.123, 3.142, and 6.074, respectively, df = 4). (e, f) Expression of GRB10 in neurons treated with KA, IGF-1, ADAR1, and their inhibitors ( t = 11.242, 2.162, and 7.230, respectively, df = 4). ∗ , #, & P < 0.05.

Article Snippet: Goat β -actin antibody (ab8227) and rabbit anti-ADAR1 antibody (bs-2168R; Bioss, Woburn, MA, USA), rabbit polyclonal to GRB10 (ab125583; Abcam, Cambridge, UK), mouse monoclonal to Bcl-2 (ab692; Abcam), and rabbit monoclonal to anti-cleaved caspase-3 (EPR21032; Abcam) were used to evaluate apoptosis in mice with TBI.

Techniques: Immunoprecipitation, Expressing

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: RNA isolated from brains of 4 to 5 week old WT or RdRp tg/- mice (n = 5 for WT, 6 for RdRp tg/- ) was used to measure (A) ADAR p110 and (B) ADAR p150 transcripts by qPCR. Data points represent individual animals, graphs shown means, s.d. (C) Immunoblotting for ADAR p110 and ADAR p150 in 4 to 5 week old mouse WT and RdRp tg/- brain. **** = p < 0.0001; n = 3 animals per genotype. (D) A549 cells with inducible Theiler’s virus RdRp (Tet-on system). Data are shown from two representative knockdowns. (E) RNAs isolated from the parallel knockdowns done in panel D were used for qPCR analysis to determine relative levels of the ISGs OAS and ISG15 mRNAs. mRNAs were harvested 54 hours after ADAR -targeting siRNA addition and 48 hours after dox addition. Comparisons were made between dox-treated and dox-untreated cells and expressed as fold mRNA changes induced by dox. Transfection controls (cells receiving transfection reagents but no siRNA) showed similar levels of ISG induction after dox treatment as control siRNA-transfected cells, indicating lack of contribution of transfected siRNAs to immune activation. Data are means and s.d. of triplicate biological replicates with three technical replicates each. NS: not significant; unpaired Student’s T test.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Isolation, Western Blot, Virus, Transfection, Control, Activation Assay

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: (A) Size and coat color differences in 5-week-old, littermate RdRp tg/- and RdRp tg/- Adar +/- mice. (B) Weight differences between 4 week and 5-week-old littermate RdRp tg/- and RdRp tg/- Adar +/- mice (n = 9 and 5 respectively). (C) Dental formation changes between RdRp tg/- and RdRp tg/- Adar +/- mice. (D) H & E-stained tissue sections from kidney, brain, lung, liver, heart and spleens from RdRp tg/- and RdRp tg/- Adar +/- littermate mice. Pathological grading revealed no significant scoring in RdRp tg/- Adar +/- animals. (E) Blood Urea nitrogen from serum from 4-5 week old animals in WT, Adar +/- , RdRp tg/- , Adar +/- RdRp tg/- mice. n = 5, 6, 5, 8 for WT, Adar +/- , RdRp tg/- , Adar +/- RdRp tg/- , respectively. (F) Anti-SmAG antibodies from serum from 4-5 week old animals as measured by quantitative ELISA. Four outlier high values were seen, but two were in the WT group. n = 15, 7, 10, 9 for WT, Adar +/- , RdRp tg/- , Adar +/- RdRp tg/- , respectively. Anti-dsDNA antibody levels from matched serum was below the limit of detection for all animals tested. Where not indicated, all data and tissue sections come from a mix of male and female mice. Data in (B) was analyzed by two-way ANOVA where ****p < 0.0001. Data in (E) and (F) were analyzed by one-way ANOVA where *p < 0.05; data points represent individual animals, graphs show means and s.d.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Femurs from 6-week-old animals were manually de-fleshed and used for all analyses. (A) Femur lengths of WT, Adar +/- , RdRp tg/- , and RdRp tg/- Adar +/- mice (n = 8, 6, 10, and 17 animals respectively). (B) Representative µCT images, distal femur sections and whole bone. (C) Cortical bone volume fraction (BV/TV), n = 8, 7, 10, 17. (D) Trabecular number, n = 8, 7, 9, 17. (E) Trabecular separation, n = 8, 7, 9, 17. (F) Stiffness, n = 8, 7, 10, 15. (G) Maximum load, n = 8, 7, 10, 17. (H) Cortical total mineral density, n = 8, 7, 10, 17. (I) Modulus, n = 8, 7, 10, 17. (J) Ultimate stress, n = 8, 7, 10, 17. Mice with gray fur are indicated by light-blue symbols. Data were analyzed first by using a ROUT test to remove outliers (Q = 1%), which resulted in removal of two mice from one group in one panel (the RdRp tg/- Adar +/- group in , stiffness testing; hence there are 15 mice as opposed to the 17 for this genotype in the other panels). A one-way ANOVA comparing RdRp tg/- Adar +/- to each other group was used followed by a Tukey tests where * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. Data points represent individual animals, horizontal bars indicate means.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques:

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Flash frozen brain and formalin-fixed paraffin embedded heart were collected from 6-week-old mice. (A) Heart samples were stained with Alizarin Red and calcium depositions were manually counted and shown as Alizarin Red foci per animal (n = 3, 5, 5, 10 for WT, Adar +/- , RdRp tg/ , RdRp tg/- Adar +/- , respectively). Representative heart section images are shown to the right: (i) positive control heart provided by UCDSOM Research Histology Core Laboratory; (ii) RdRp tg/- Adar +/- heart without Alizarin red foci; (iii) RdRp tg/- Adar +/- heart with a positive focus. (B) Calcium was quantified from brain tissues (n = 3, 4, 5, 5). Data were analyzed using a one-way ANOVA followed by a Tukey tests; there were no significant differences between the groups. ns: not significant.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Formalin-fixed Paraffin-Embedded, Staining, Positive Control

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: RNA-seq from neuronal tissue from n = 4 age-matched (5 weeks) mice per group. Where possible, littermate controls were used for analysis. Two male and two female mice were used per group. (A) Multi-dimensional scaling plot of RNA expression profiles in WT-WT, Adar +/- , RdRp tg/- , and RdRp tg/- Adar +/- mice. (B) Heatmap of differentially expressed ISGs across the four genotypes. FPKM values were log transformed with 1 pseudocount to facilitate visualization. (C) Summary table of differentially expressed genes (DEGs) and proportions of which are known ISGs as determined by the Interferome database. (D) Differentially expressed genes that are shared across all group comparisons. (E) Canonical molecular pathways from IPA that are significant across group comparisons.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: RNA Sequencing Assay, RNA Expression, Transformation Assay

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Predicted upstream signaling regulators as determined by IPA in various genotype comparisons between WT, Adar +/- , RdRp tg/- , and Adar +/- RdRp tg/- mice where red/green indicates whether the gene itself is upregulated/downregulated in the RNA-seq and orange/blue indicates predicated activation/inhibition of the molecular pathway it regulates.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: RNA Sequencing Assay, Activation Assay, Inhibition

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: (A) qPCR of three representative ISGs ( Ifit1 , Isg15 , and Oasl2 ) in brain tissue of 4-5 week old WT, Adar +/- , RdRp tg/ , and RdRp tg/- Adar +/- mice (n = 5, 5, 6, 5, respectively). (B-E) Flow cytometry analysis of cellular populations derived from the spleens of 4-5 week old mice. Single cell, RBC-lysed solutions were prepared for use in analysis. (B) BST-2 expression on the main immune cell subsets in the spleen including T cells (CD3 + ), B cells (CD19 + ), DCs (CD11c + ), granulocytes (Ly6G + ), and NK cells (NKp46 + ). n = 7, 7, 4, 4 for WT, Adar +/- , RdRp tg/ , RdRp tg/- Adar +/- , respectively. (C) Monocytes populations across all four groups. (D) BST-2 expression on monocyte/DC cell subsets including monocytes, cDC1s, cDC2s, and moDCs. (D) Activation (CD80/86 expression) of monocyte/DC subsets including monocytes, cDC1s, cDC2s, and moDCs. C-E: n = 12, 7, 10, 8 for WT, Adar +/- , RdRp tg/ , RdRp tg/- Adar +/- , respectively. For all graphs shown, data were analyzed using a one-way ANOVA followed by a Tukey tests to determine significance where * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. Data points represent individual animals with the mean and s.d. shown as bars.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Flow Cytometry, Derivative Assay, Expressing, Activation Assay

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: 6-week-old animals were weighed (A) and RNA was isolated from whole-brains for qPCR analysis of the ISGs Ifit1 (B) , Isg15 (C) , and Oasl2 (D) . Data were analyzed using a one-way ANOVA followed by a Tukey tests where * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. For (A), n = 4, 5, 6, 10, and 4 for WT, Adar +/- , RdRp tg/ , RdRp tg/- Adar +/- , and RdRp tg/- Adar +/- Ifih1 -/- , respectively. For (B-D), n = 4, 3, 5, 5, and 4. Data points represent individual animals with the mean for each group shown as a bar.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Isolation

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Enriched diseases and biological functions across comparisons of gene expression changed between WT, Adar +/- , RdRp tg/- , and Adar +/- RdRp tg/- mice. Orange indicated that the disease or biological function is predicted to be enriched for and blue indicated is predicted to be inhibited.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Expressing

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Splenocytes from 4-5 week old mice were harvested and single cell, RBC-lsysed solutions were made for FACS analysis. (A) T cells, B cells, DCs, granulocytes, and NK cells show no difference across groups measured. n = 7, 7, 4, and 4 for WT, Adar +/- , RdRp tg/ , and RdRp tg/- Adar +/- , respectively. (B) DC subsets including pDCs, cDC1s, cDC2s, and moDCs show minor or no difference in populations across groups. n = 12, 7, 10, and 8 for WT, Adar +/- , RdRp tg/ , and RdRp tg/- Adar +/- , respectively. All data was analyzed using a one-way ANOVA followed by a Tukey test to determine significance. * = p < 0.05. Data represent individual animals, graphs show means and s.d.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques:

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Changes in A-to-I edits were determined via comparison of RNA-seq data with whole exome DNA sequencing. (A) Pie charts showing the proportion of editing sites within a given RNA element for each mouse genotype. (B) Table summary of the distribution of overall numbers of edited genes and sites for each genotype and the breakdown of locations of the edit sites among different RNA elements. For each of the four genotypes, four animals were sequenced and to be counted a gene must have been edited in all four animals sequenced but editing can occur anywhere in the gene. For sites, identical sites must be edited in all four animals of a genotype. (C) Venn diagrams showing overlap of edited genes (top) or sites (bottom) among the four genotypes. (D,E) Immunoblot analysis from two animals per group (D) and gene expression fold change (E) of three Adar +/- RdRp tg/- uniquely edited genes in the RNA-seq data set (as compared to WT) in 5-week-old neuronal tissue. Rig-I, Isg15, and Zbp1 were evaluated as they are each ISGs that are highly upregulated in mice expressing RdRp and are key regulators of the antiviral response. (F) Expression of Adar isoforms in neuronal tissue of 4-5 week old mice from all four genotypes, as determined by western blot (n = 2 per group).

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: Comparison, RNA Sequencing Assay, DNA Sequencing, Western Blot, Expressing

Journal: bioRxiv

Article Title: ADAR1 haploinsufficiency and sustained viral RdRp dsRNA synthesis synergize to dysregulate RNA editing and cause multi-system interferonopathy

doi: 10.1101/2025.01.21.634124

Figure Lengend Snippet: Genes with A-to-I edit sites and RNA-seq-derived expression changes were analyzed using IPA to determine enriched diseases and biological functions across genotype comparisons (WT, Adar +/- , RdRp tg/- , and Adar +/- RdRp tg/- ). Orange indicates that the disease or biological function is predicted to be enriched for or activated and blue indicates is predicted to be inhibited.

Article Snippet: For human cell line derived blots, rabbit polyclonal anti-ADAR1 (1:1,000 dilution; Cell Signaling, 14175) primary antibody or mouse anti-tubulin (1:5,000 dilution; Sigma, T5168) were used.

Techniques: RNA Sequencing Assay, Derivative Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Targeting exosomal double-stranded RNA-TLR3 signaling pathway attenuates morphine tolerance and hyperalgesia

doi: 10.1016/j.xcrm.2024.101782

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ADAR1 , Proteintech , Cat#14330-1-AP; RRID: AB_2273600.

Techniques: Virus, Recombinant, CCK-8 Assay, Modification, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Transfection, Sequencing, Negative Control, Software