Journal: Viruses

Article Title: Activation of Interferon-Stimulated Genes following Varicella-Zoster Virus Infection in a Human iPSC-Derived Neuronal In Vitro Model Depends on Exogenous Interferon-α

doi: 10.3390/v14112517

Figure Lengend Snippet: Human interferon pathway: Assay IDs and targets.

Article Snippet: Hs00194264_m1 , IFNGR2.

Techniques:

Journal: European journal of immunology

Article Title: Influenza A virus abrogates IFN-gamma response in respiratory epithelial cells by disruption of the Jak/Stat pathway.

doi: 10.1002/eji.200737045

Figure Lengend Snippet: Figure 11. Detection of IFNGR1, IFNGR2, Jak1, Jak2, and Stat1a expression in influenza A virus-infected A549 cells. A549 cells were infected with influenza Aichi at an MOI of 10 or left uninfected as described in Fig. 2. The infected cells and uninfected control cells were treated with IFN-c (1000 U/mL) for 1 h or left untreated. (A) Total RNA was extracted and subject to RT-PCR analysis with primers specific for IFNGR1, IFNGR2, Jak1, Jak2, and Stat1. (B) The protein levels were detected by western blot assay. A549 cells were infected and treated with IFN-c as described above. Total cell lysates (40 lg) were directly analyzed by western blotting with Ab to IFNGR1, IFNGR2, Jak1, Jak2, and Stat1a. Results shown are representative of three separate experiments.

Article Snippet: For detection of IFNGR1, IFNGR2, Jak1, and Jak2 , primary Ab used for western analyses included a rabbit polyclonal affinity purified anti-IFNGR1 Ab (1:2000; Santa Cruz Biotechnology), a rabbit polyclonal affinity purified anti-IFNGR2 Ab (1:1000; Protein Tech Group Inc., Chicago, IL), a rabbit polyclonal antihuman Jak1 Ab (1:5000; Upstate Biotechnology), and a rabbit polyclonal affinity purified anti-Jak2 Ab (1:1000; Santa Cruz Biotechnology).

Techniques: Expressing, Virus, Infection, Control, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Nature

Article Title: Autoimmunity in Down’s syndrome via cytokines, CD4 T cells and CD11c + B cells

doi: 10.1038/s41586-023-05736-y

Figure Lengend Snippet: a , PCA analysis of the HuProt IgG dataset for adults with DS ( n = 5), age-matched HC individuals ( n = 4), and patients with IPEX ( n = 3) and APS-1 ( n = 1). b , The number of IgG autoantigens enriched at least twofold in individuals with DS, IPEX and APS-1 compared with HC individuals. c , Enriched IgG autoantigens overlapping between disease groups. d , Enriched IgG autoantigens in HC individuals and in individuals with DS, IPEX and APS-1. The colour intensity corresponds to the log 2 -transformed fold change expression value relative to the mean of healthy adult controls. F, female; M, male; NA, unknown. e , Chromosomal expression pattern of IgG autoantigens enriched in DS. f , Gene expression pattern of IgG autoantigens enriched in DS ( n = 5) according to the Human Protein Atlas. g , GO analysis of IgG autoantigens enriched in DS ranked by the number of autoantigens found to be enriched in the associated gene set. The dot size and colour intensity correspond to the FDR-adjusted P value. Med., mediated; NK, natural killer; reg., regulation; sig, signalling; surf, surface; sys., system. h , The surface expression of CD64 in monocytes, natural killer cells from individuals with DS ( n = 14) and age-matched HC individuals ( n = 8). Mono, monocytes. i , ELISA analysis of anti-IFNGR2 autoantibodies in the plasma from adults with DS ( n = 4) and age-matched HC individuals ( n = 7). j , Neutralizing IFNγ signalling in THP-1 cells by IgG fraction of plasma from adults with DS ( n = 3), age-matched HC individuals ( n = 3) or recombinant anti-IFNGR2 antibody. Significance was assessed using one-way ANOVA with Tukey’s post hoc analysis. For h and i , significance was assessed using two-tailed unpaired t -tests. For h and j , data are mean ± s.d. For i , the whiskers denote the minimum and maximum values, the box limits denote quartile 1 to quartile 3, and the centre bar denotes the mean.

Article Snippet: Overnight, 96-well Costar plates were coated at 4 °C with 100 μl per well of a 1 μg ml −1 solution of recombinant IFNGR2, MSTN or ATP6V1G2 protein (OriGene) suspended in 1× PBS.

Techniques: Immunopeptidomics, Transformation Assay, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Recombinant, Two Tailed Test

Journal: Brain, behavior, and immunity

Article Title: Antinociceptive interactions between excitatory interferon-γ and interleukin-17 in sensory neurons.

doi: 10.1016/j.bbi.2024.11.021

Figure Lengend Snippet: Figure 3. IFNγ receptor expression in adult rat dorsal root ganglion (DRG) sections. (A) Labeling of DRG neuronal profiles with antibodies against IFNGR1 or IFNGR2 and controls. Arrows indicate typical neurons labeled with the antibodies. In the control section the primary antibody has been omitted. The right image shows a negative isotype-control. (B) Left: Fluorescence intensity of neurons from control incubations (red)

Article Snippet: We used a rabbit polyclonal antibody against the C-terminus of IFNGR1 (#ABIN686902, antibodies-online, Aachen, Germany), diluted 1:200, and a rabbit polyclonal antibody against the C-terminus of IFNGR2 (#orb5501, Biorbyt, Cambridge, UK), diluted 1:100.

Techniques: Expressing, Labeling, Control, Fluorescence

Journal: Immunity

Article Title: T-bet transcription factor promotes antibody secreting cell differentiation by limiting the inflammatory effects of IFNγ on B cells

doi: 10.1016/j.immuni.2019.04.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: TaqMan Gene Expression Assay Ifngr2 , Applied Biosystems , Mm01210592_m1.

Techniques: Blocking Assay, Virus, Recombinant, Purification, Gene Expression, Staining, Reverse Transcription, DNA Library Preparation, cDNA Synthesis, Labeling, Microarray, Software

Journal: Cell Death Discovery

Article Title: Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon- γ -induced malignant transformation of primary bovine mammary epithelial cells

doi: 10.1038/cddiscovery.2015.65

Figure Lengend Snippet: Expression of IFNGRs in cow mammary glands. Two groups of Holstein cows were fed with mixed forage (MF) or corn straw (CS). The experimental period was 12 weeks, and the pre-feeding period was 3 weeks. At the end of the feeding trial, the expression levels of IFNGR1 and IFNGR2 in mammary tissue (obtained via biopsy) were analysed using immunohistochemical staining and western blot analysis. ( a ) Immunohistochemical staining of IFNGR1 and IFNGR2. Scale bars, 100 μ m. Insets (scale bars, 200 μ m) show the overall presence of the brown colour indicating IFNGR1 and IFNGR2. Statistical analysis of the grey colour intensity (right). The data represent the mean±S.E.M. of three independent experiments. Error bars are±S.E.M. One-way ANOVA; * P <0.05. ( b ) Detection of IFNGRs via western blot analysis as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05.

Article Snippet: The main primary antibodies used in this study were specific for MAP1LC3 (Cell Signaling Technology, Beverly, MA, USA; Cat 2775), ATG5 (Novus Biologicals, Littleton, CO, USA; Cat NB110-53818), SQSTM1/p62 (Santa Cruz Biotechnology Inc., Heidelberg, Germany; Cat sc-25575), DAPDH (Cell Signaling Technology; Cat 2118), IFNGR1 (Bioss Biotechnology Co., LTD., Beijing, China; Cat bs-1463R), IFNGR2 (Bioss Biotechnology Co., LTD.; Cat bs-2710R), LC3B (Bioss Biotechnology Co., LTD; Cat bs-4843R) and SQSTM1 (Bioss Biotechnology Co., LTD; Cat bs-2951R).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Cell Death Discovery

Article Title: Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon- γ -induced malignant transformation of primary bovine mammary epithelial cells

doi: 10.1038/cddiscovery.2015.65

Figure Lengend Snippet: IFN- γ increased rates of autophagy in primary BMECs in vitro . ( a ) The expression of IFNGRs in BMECs. BMECs were treated with IFN- γ for 6, 24 or 48 h. At the end of the treatment period, the levels of IFNGR1 and IFNGR2 were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( b ) BMECs were treated with IFN- γ for 3, 6, 12 or 24 h. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( c ) BMECs were treated with IFN- γ at 2.5, 5, 10 or 20 ng/ml. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( d ) Direct effect of IFN- γ on induction of autophagy. BMECs were cultured for 24 h in supernatants from either mock-treated BMECs or cells treated with 10 ng/ml of IFN- γ for 24 h in the presence or absence of anti-IFNGR1, IFNGR2 or IFNGR1/2. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01.

Article Snippet: The main primary antibodies used in this study were specific for MAP1LC3 (Cell Signaling Technology, Beverly, MA, USA; Cat 2775), ATG5 (Novus Biologicals, Littleton, CO, USA; Cat NB110-53818), SQSTM1/p62 (Santa Cruz Biotechnology Inc., Heidelberg, Germany; Cat sc-25575), DAPDH (Cell Signaling Technology; Cat 2118), IFNGR1 (Bioss Biotechnology Co., LTD., Beijing, China; Cat bs-1463R), IFNGR2 (Bioss Biotechnology Co., LTD.; Cat bs-2710R), LC3B (Bioss Biotechnology Co., LTD; Cat bs-4843R) and SQSTM1 (Bioss Biotechnology Co., LTD; Cat bs-2951R).

Techniques: In Vitro, Expressing, Western Blot, Cell Culture