abs against stat1  (Santa Cruz Biotechnology)

Journal: Transboundary and Emerging Diseases

Article Title: Novel Duck Orthoreovirus σA Protein Inhibits Interferon Signaling by Impeding STAT1/STAT2 Nuclear Translocation

doi: 10.1155/tbed/8440800

Figure Lengend Snippet: σA does not affect the expression or phosphorylation of proteins related to JAK-STAT signaling pathway, nor does it interact directly with STAT1/STAT2. (A) HEK293T cells were transfected with HA-σA or an empty vector. After 24 h, cells were treated with 500 U/mL IFN-α. Total RNA and protein were extracted, and the expression levels of IFNAR1, IRF9, and JAK1 at the transcriptional and protein levels were analyzed by qPCR and western blot, respectively. (B) Under the same conditions, the transcriptional activity, expression, and phosphorylation of STAT1 and STAT2 were assessed by qPCR and western blot. (C) HEK293T cells were transfected with HA-σA or an empty vector. After 24 h, cells were treated with 500 U/mL IFN-α. Total protein lysates were immunoprecipitated with anti-HA antibody and analyzed by western blot using anti-STAT antibodies.

Article Snippet: Abs against STAT1, STAT2, and β-tubulin were purchased from Proteintech (Catalog Numbers 10144, 16674, and 10094, USA).

Techniques: Expressing, Phospho-proteomics, Transfection, Plasmid Preparation, Western Blot, Activity Assay, Immunoprecipitation

Journal: Transboundary and Emerging Diseases

Article Title: Novel Duck Orthoreovirus σA Protein Inhibits Interferon Signaling by Impeding STAT1/STAT2 Nuclear Translocation

doi: 10.1155/tbed/8440800

Figure Lengend Snippet: σA protein inhibits IFN-α-induced STAT1/STAT2 nuclear translocation. (A) HEK293T cells were transfected with HA-σA plasmid or empty vector. After 24 h, cells were treated with 500 U/mL IFN-α, and the subcellular localization of STAT1/STAT2 was analyzed by confocal microscopy. (B) HEK293T cells were transfected with HA-σA plasmid or empty vector. After 24 h, cells were treated with 500 U/mL IFN-α. Nuclear and cytoplasmic fractions were isolated, and STAT1/STAT2 phosphorylation was assessed by western blot.

Article Snippet: Abs against STAT1, STAT2, and β-tubulin were purchased from Proteintech (Catalog Numbers 10144, 16674, and 10094, USA).

Techniques: Translocation Assay, Transfection, Plasmid Preparation, Confocal Microscopy, Isolation, Phospho-proteomics, Western Blot

Journal: Transboundary and Emerging Diseases

Article Title: Novel Duck Orthoreovirus σA Protein Inhibits Interferon Signaling by Impeding STAT1/STAT2 Nuclear Translocation

doi: 10.1155/tbed/8440800

Figure Lengend Snippet: Schematic diagram illustrating the mechanism through which the NDRV σA protein inhibits the nuclear translocation of STAT1/STAT2. The σA protein significantly suppresses the IFN signaling pathway and its associated antiviral effects by specifically blocking the nuclear translocation of STAT1/STAT2, thereby, enabling the virus to evade the host's innate immune response.

Article Snippet: Abs against STAT1, STAT2, and β-tubulin were purchased from Proteintech (Catalog Numbers 10144, 16674, and 10094, USA).

Techniques: Translocation Assay, Blocking Assay, Virus

Journal: Mediators of Inflammation

Article Title: The Spliceosome Factor EFTUD2 Promotes IFN Anti-HBV Effect through mRNA Splicing

doi: 10.1155/2023/2546278

Figure Lengend Snippet: EFTUD2 regulation of classical ISGs did not act through direct regulation of the JAK-STAT pathway. (a–d) RT-qPCR detected the STAT1, IRF9, IFNAR1, and JAK1 mRNA in WT and EFTUD2 +/- HepG2.2.15 cells before and after IFN treatment. (e) After incubated with 5000 IU/mL IFN- α for 24 h, STAT1 and STAT1 phosphorylation (P-STAT1) were assessed by Western blotting in WT and EFTUD2 +/- HepG2.2.15 cells.

Article Snippet: And Abs against STAT1 (diluted at 1 : 1000) and p-STAT1 (diluted at 1 : 1000) were purchased from Cell Signaling.

Techniques: Quantitative RT-PCR, Incubation, Phospho-proteomics, Western Blot

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

Article Title: SOCS1 Is a Key Molecule That Prevents Regulatory T Cell Plasticity under Inflammatory Conditions.

doi: 10.4049/jimmunol.1600441

Figure Lengend Snippet: FIGURE 6. Increased production of IL-12p40 by APCs affected by Socs1-deficient Tregs and activation of STAT4 in Socs1-deficient Tregs by IL-12 for conversion into ex-Tregs. (A) APCs (2 3 105 cells per well) from Socs1+/+ or Socs1fl/flFoxp3YFP-Cre mice (,12 wk) mice were cultured for 72 h with IFN-g (50 ng/ml) or CpG (1 mM), following which IL-12p40 levels in the culture supernatant were measured using ELISA. Data are presented as the mean 6 SD of three independent experiments. Cultured APCs were sorted from six animals in each group. **p , 0.01, Student t test and Kruskal–Wallis tests with the Bonferroni test. (B) Tregs (5 3 104 cells per well) and APCs (2 3 105 cells per well) from WT mice were cocultured for 4 d (WT). Similarly, Tregs and APCs from Socs1fl/flLck-Cre+ mice were cocultured for 4 d (Socs1fl/flLck-Cre+). IL-12p40 levels in the culture supernatant were measured using ELISA. Data are presented as the mean 6 SD of three independent experiments. **p , 0.01, Student t test. Tregs were sorted from 12 animals in each group, and APCs were sorted from three animals in each group. (C) Analysis of the phosphorylation of STAT1, STAT3, and STAT4 in Tregs. Tregs (1 3 105 cells) from WT or Socs1fl/flFoxp3YFP-Cre mice were stimulated with or without IL-12 (40 ng/ml) for 30 min, following which phosphoSTATs were detected by immunoblotting. Tubulin was analyzed as a loading control. Data are representative of three independent experiments. Cultured Tregs were sorted from 12 animals in each group.

Article Snippet: Abs against phospho-STAT1, phospho-STAT3, and phospho-STAT4 (all from Cell Signaling Technology) and tubulin (Sigma) were used to visualize the corresponding proteins.

Techniques: Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Western Blot, Control