Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatitis delta virus inhibits alpha interferon signaling.

doi: 10.1002/hep.22654

Figure Lengend Snippet: Fig. 3. The nuclear accumulation of STAT1 and STAT2 in response to IFN- is inhibited in HDV-transfected cells. Normal cells (Huh-7) or cells transfected with HDV cDNA [Huh-7 pS- VL(D3)] were seeded on glass coverslips and treated with 1000 U/mL IFN- for 30 minutes. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, and stained with antisera to STAT1 (A) or STAT2 (B) and HDAg. Nuclei were counterstained with 4,6-diamidino-2-phenylindole (blue). Arrows point to cells successfully transfected with HDV.

Article Snippet: Rabbit polyclonal antibodies to STAT1, STAT2, Jak1, Tyk2, phosphotyrosine 701- STAT1 (pY-STAT1), phosphotyrosine 690-STAT2 (pYSTAT2), phosphotyrosine 1022/1023-Jak1 (pY-Jak1), and phosphotyrosine 1054/1055-Tyk2 (pY-Tyk2) were from Cell Signaling Technology, Inc. (Danvers, MA).

Techniques: Transfection, Staining

Journal: Hepatology (Baltimore, Md.)

Article Title: Hepatitis delta virus inhibits alpha interferon signaling.

doi: 10.1002/hep.22654

Figure Lengend Snippet: Fig. 4. HDV inhibition of IFN-–induced tyrosine phosphorylation of STAT1 and STAT2. Normal cells (Huh-7) and cells transfected with HDV cDNA [Huh-7 pSVL(D3)] were cultured for 9 days and then treated with 1000 U/mL IFN- for 30 minutes or left untreated. Equal amounts of cell lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, transferred onto nitrocellulose membranes, and exam- ined with antibodies to phosphotyrosine 701-STAT1 (pY-STAT1), STAT1, phosphotyrosine 690-STAT2 (pY-STAT2), STAT2, p48, and HDAg. -Actin levels served as a loading control.

Article Snippet: Rabbit polyclonal antibodies to STAT1, STAT2, Jak1, Tyk2, phosphotyrosine 701- STAT1 (pY-STAT1), phosphotyrosine 690-STAT2 (pYSTAT2), phosphotyrosine 1022/1023-Jak1 (pY-Jak1), and phosphotyrosine 1054/1055-Tyk2 (pY-Tyk2) were from Cell Signaling Technology, Inc. (Danvers, MA).

Techniques: Inhibition, Phospho-proteomics, Transfection, Cell Culture, Polyacrylamide Gel Electrophoresis, Control

Journal: Journal of Biological Chemistry

Article Title: Viral Induction of the Zinc Finger Antiviral Protein Is IRF3-dependent but NF-κB-independent

doi: 10.1074/jbc.m109.054486

Figure Lengend Snippet: FIGURE 7. Differential binding of IRF3 and STAT transcription factors to the hZAP promoter following virus infection or IFN stimulation. A, immu- noblot analysis of ISG56 expression and phosphorylation status of STAT1 and STAT2 in HeLa cells following stimulation of IFN (500 units/ml) for 1 and 8 h or infection with SeV (100 HAU/ml) for 16 h. Actin blot was shown as a loading control. B, ChIP analyses of IRF3 and STAT binding to the ISRE1–2, ISRE3–5, and STAT V sites within hZAP promoter in HeLa cells mock treated, stimulated with IFN (400 units/ml) for 1 h, or infected with SeV (200 HAU/ml) for 8 h. The ChIP-enriched DNA levels analyzed by Q-PCR were normalized to input DNA, followed by subtraction of nonspecific binding determined with control IgG.

Article Snippet: Sheared chromatin was incubated with 2 g of control IgG (Active Motif) or specific antibody to IRF3, STAT1, or STAT2 (Santa Cruz), and 25 l of protein G magnetic beads overnight at 4 °C.

Techniques: Binding Assay, Virus, Infection, Expressing, Phospho-proteomics, Control

Journal: Current Oncology

Article Title: STAT2 Promotes Tumor Growth in Colorectal Cancer Independent of Type I IFN Receptor Signaling

doi: 10.3390/curroncol32120707

Figure Lengend Snippet: STAT2 expression is elevated in colon tumors and correlates with reduced survival. Boxplots showing ( a ) STAT2 and ( c ) IFNAR1 mRNA expression are significantly higher in TCGA-COAD tumor tissues ( n = 286) compared with normal colon samples ( n = 41). ( b ) Kaplan–Meier survival curves for patients stratified by median STAT2 expression show reduced overall survival in the STAT2-high group. ( d ) STAT2-high tumors stratified by IFNAR1-high ( n = 56) and IFNAR-low ( n = 54) expression show no survival difference. Cox proportional hazards model was applied. Statistical significance; p ≤ 0.05.

Article Snippet: Anti-STAT1 antibody (Cat#10144-2-AP), anti-STAT2 antibody (cat##51075-2-AP), anti-STAT3 (cat#10253-2-AP), anti-β-tubulin (cat#10094-1-AP), and anti-β-Actin (cat#66009-1-Ig) were purchased from Proteintech (Rosemont, IL, USA.

Techniques: Expressing

Journal: Current Oncology

Article Title: STAT2 Promotes Tumor Growth in Colorectal Cancer Independent of Type I IFN Receptor Signaling

doi: 10.3390/curroncol32120707

Figure Lengend Snippet: STAT3 activation is preserved in STAT2 KO tumor cells. ( a ) Western blots confirming basal levels of STAT1, STAT2, and STAT3 after deletion of STAT2 or IFNAR1 in knockout clones. ( b ) Time course analyses of phosphorylated STATs following IFN-I stimulation. ( c ) Impaired transcriptional response to IFN-I in both STAT2 KO and IFNAR1 KO cells. *** p ≤ 0.001; **** p ≤ 0.0001. ns, not statistically significant.

Article Snippet: Anti-STAT1 antibody (Cat#10144-2-AP), anti-STAT2 antibody (cat##51075-2-AP), anti-STAT3 (cat#10253-2-AP), anti-β-tubulin (cat#10094-1-AP), and anti-β-Actin (cat#66009-1-Ig) were purchased from Proteintech (Rosemont, IL, USA.

Techniques: Activation Assay, Western Blot, Knock-Out, Clone Assay

Journal: Current Oncology

Article Title: STAT2 Promotes Tumor Growth in Colorectal Cancer Independent of Type I IFN Receptor Signaling

doi: 10.3390/curroncol32120707

Figure Lengend Snippet: STAT2 and IFNAR1 differentially regulate colon cancer cell proliferation and tumor growth. ( a ) In vitro MTS showing reduced proliferation of STAT2 KO compared with parental and IFNAR1 KO HCT116 cells over the course of 72 h. Data represent mean ± SEM from n = 3. ( b ) Growth of tumor xenografts in immunodeficient Rag1KO mice injected subcutaneously with parental, STAT2 KO, or IFNAR1 KO cells. ( c ) Overexpression of STAT2 in HCT116 cells enhanced tumor growth in vivo ( n = 5–8 mice per study). * p ≤ 0.05; **** p ≤ 0.0001. ns, not statistically significant. Results are from two combined independent experiments.

Article Snippet: Anti-STAT1 antibody (Cat#10144-2-AP), anti-STAT2 antibody (cat##51075-2-AP), anti-STAT3 (cat#10253-2-AP), anti-β-tubulin (cat#10094-1-AP), and anti-β-Actin (cat#66009-1-Ig) were purchased from Proteintech (Rosemont, IL, USA.

Techniques: In Vitro, Injection, Over Expression, In Vivo

Journal: Current Oncology

Article Title: STAT2 Promotes Tumor Growth in Colorectal Cancer Independent of Type I IFN Receptor Signaling

doi: 10.3390/curroncol32120707

Figure Lengend Snippet: STAT2 signaling and IFN-I responsiveness in STAT2- and IFNAR1-deficient murine colon carcinoma cells. ( a ) Western blot analyses show IFN-I-stimulated phosphorylation of STAT1 and basal STAT1 expression in parental, STAT2 KO, and IFNAR1 KO MC38 cells. ( b ) IFN-I-stimulated phosphorylation of STAT3 and basal STAT2 expression analyzed by western blot analysis. ( c ) Impaired transcriptional responses in both KO cell lines after 6 and 24 h of IFN-β treatment. Data are shown as fold change from corresponding untreated cell genotype. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001. ns, not statistically significant.

Article Snippet: Anti-STAT1 antibody (Cat#10144-2-AP), anti-STAT2 antibody (cat##51075-2-AP), anti-STAT3 (cat#10253-2-AP), anti-β-tubulin (cat#10094-1-AP), and anti-β-Actin (cat#66009-1-Ig) were purchased from Proteintech (Rosemont, IL, USA.

Techniques: Western Blot, Phospho-proteomics, Expressing

Journal: Current Oncology

Article Title: STAT2 Promotes Tumor Growth in Colorectal Cancer Independent of Type I IFN Receptor Signaling

doi: 10.3390/curroncol32120707

Figure Lengend Snippet: Loss of STAT2, but not IFNAR1, reduces proliferation and tumor growth in murine colon carcinoma cells. ( a ) In vitro MTS assay showing reduced proliferation of Stat2 KO tumor cells compared with parental and IFNAR1 KO cell lines over 72 h. Data represent mean ± SEM from n = 3. ( b , c ) Tumor growth curves of wild-type mice injected subcutaneously with parental, STAT2 KO, or IFNAR1 KO tumor cells. * p < 0.05; *** p < 0.001; **** p < 0.0001. Data are shown as mean ± SEM from n = 6–8 mice per group.

Article Snippet: Anti-STAT1 antibody (Cat#10144-2-AP), anti-STAT2 antibody (cat##51075-2-AP), anti-STAT3 (cat#10253-2-AP), anti-β-tubulin (cat#10094-1-AP), and anti-β-Actin (cat#66009-1-Ig) were purchased from Proteintech (Rosemont, IL, USA.

Techniques: In Vitro, MTS Assay, Injection

Journal: Immunology

Article Title: Interferon-α promotes MHC I antigen presentation of islet β cells through STAT1-IRF7 pathway in type 1 diabetes.

doi: 10.1111/imm.13468

Figure Lengend Snippet: FIGURE 6 Inhibition of STAT1 and STAT2 prevented IFN-α induced MHC-I and antigen presentation in MIN6 cells. MIN6 cells were transfected with siRNA (si-STAT1 or/and si-STAT2) and then treated with or without IFN-α (1000 U/ml) for 48 h. (a) Protein and (b) mRNA of antigen presented molecules was measured. (c) Representative images (10×) of MHC I in MIN6 cells after IFN-α and siRNA treatment. *p < 0·05, **p < 0·01 and ***p < 0·001

Article Snippet: Phospho- STAT1, STAT1, Phospho- STAT2, STAT2, TAP1 and β- actin antibodies were purchased from Cell Signaling Technology company, USA; IRF7 antibody was purchased from NOVUS company, USA; B2M, PSMB8 and NLRC5 antibodies were purchased from Abcam company, UK; PhosphoSTAT3, STAT3 and Phospho- STAT5 antibodies were purchased from Abclonal company, China; STAT5 and IRF9 antibodies were purchased from company, China.

Techniques: Inhibition, Immunopeptidomics, Transfection

Journal: Immunology

Article Title: Interferon-α promotes MHC I antigen presentation of islet β cells through STAT1-IRF7 pathway in type 1 diabetes.

doi: 10.1111/imm.13468

Figure Lengend Snippet: FIGURE 8 IFN-α induce β cells autoimmunity by promoting MHC I antigen presentation. In β cells, IFN-α binds to the IFNRs and lead to the nuclear translocation of STAT1 and IRF7, which in turn activates the STAT1-IRF7 axis to increase expression of antigen presenting molecules (TAP1, PSMB8 and MHC I). These actions create positive feedback through IRF7-STAT2 cascade amplifying signals and promote the proliferation of CD8+ T cells and insulitis. Figure was created with BioRender.com (Agreement number: TX23NA2XMH)

Article Snippet: Phospho- STAT1, STAT1, Phospho- STAT2, STAT2, TAP1 and β- actin antibodies were purchased from Cell Signaling Technology company, USA; IRF7 antibody was purchased from NOVUS company, USA; B2M, PSMB8 and NLRC5 antibodies were purchased from Abcam company, UK; PhosphoSTAT3, STAT3 and Phospho- STAT5 antibodies were purchased from Abclonal company, China; STAT5 and IRF9 antibodies were purchased from company, China.

Techniques: Immunopeptidomics, Translocation Assay, Expressing