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

Journal: Scientific Reports

Article Title: STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

doi: 10.1038/srep38336

Figure Lengend Snippet: ( A ) STAT2 knockout (KO) clones #1 and #2 were established from Huh-7.5 cells expressing a STAT2 sgRNA. DNA and amino acid sequences surrounding the sgRNA target sequences (blue) are shown. The protospacer adjacent motif (PAM) and the mutation in each allele are shown in green and red, respectively. ( B ) NT sgRNA-expressing cells and STAT2 KO cells (clones #1 and #2) were infected with HCVcc and treated with IFN-α (1,000 U/ml) for the indicated times. The expression levels of NS5A, PKR, and STAT2 were evaluated by immunoblotting (IB). ( C ) NT sgRNA-expressing cells and STAT2 KO cells were infected with HCVcc and treated with IFN-α (1,000 U/ml) for 96 h. Intracellular HCV RNA levels were quantified by real-time PCR and normalized to control values. Data represent the mean ± S.D. ( n = 3). * P < 0.01. NS, not significant.

Article Snippet: Anti-phospho-STAT2 (600-401-A93S) antibody was purchased from Rockland Immunochemicals.

Techniques: Knock-Out, Clone Assay, Expressing, Mutagenesis, Infection, Western Blot, Real-time Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

doi: 10.1038/srep38336

Figure Lengend Snippet: ( A , B ) STAT3 knockout (KO) clones #1, #2 were established from Huh-7.5 cells expressing STAT3 sgRNAs #1 and #2, respectively. STAT6 knockout (KO) clones #1 and #2 were established from Huh-7.5 cells expressing STAT6 sgRNAs #1 and #2, respectively. ( A ) Cells were infected with HCVcc and treated with IFN-α (1,000 U/ml) for 72 h. The expression levels of NS5A, PKR, IRF9, STAT1, STAT2, STAT3, and STAT6 were evaluated by immunoblotting (IB). ( B ) Cells were infected with HCVcc and treated with IFN-α (1,000 U/ml) for 96 h. Intracellular HCV RNA levels were quantified by real-time PCR and normalized to control values. Data represent the mean ± S.D. ( n = 3). * P < 0.01. NS, not significant.

Article Snippet: Anti-phospho-STAT2 (600-401-A93S) antibody was purchased from Rockland Immunochemicals.

Techniques: Knock-Out, Clone Assay, Expressing, Infection, Western Blot, Real-time Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

doi: 10.1038/srep38336

Figure Lengend Snippet: ( A ) NT sgRNA-expressing Huh-7.5 cells, STAT1 knockout (KO) cells (clones #1 and #2), and STAT2 KO cells (clone #1) were infected with HCVcc and treated with IFN-λ (1,000 U/ml) for the indicated times. The expression levels of NS5A, PKR, STAT1, and STAT2 were evaluated by immunoblotting (IB). ( B ) NT sgRNA-expressing cells, STAT1 KO cells (clones #1 and #2), and STAT2 KO cells (clone #1) were infected with HCVcc and treated with IFN-λ (1,000 U/ml) for 96 h. Intracellular HCV RNA levels were quantified by real-time PCR and normalized to control values. Data represent the mean ± S.D. ( n = 3). * P < 0.01. NS, not significant.

Article Snippet: Anti-phospho-STAT2 (600-401-A93S) antibody was purchased from Rockland Immunochemicals.

Techniques: Expressing, Knock-Out, Clone Assay, Infection, Western Blot, Real-time Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

doi: 10.1038/srep38336

Figure Lengend Snippet: ( A ) NT sgRNA-expressing Huh-7.5 cells and STAT1 knockout (KO) cells (clone #2) were infected with HCVcc and treated with IFN-α (1,000 U/ml) or IFN-λ (1,000 U/ml) for the indicated times. The expression levels of NS5A, PKR, IRF9, STAT1, and STAT2 were evaluated by immunoblotting (IB). ( B ) NT sgRNA-expressing cells, STAT1 KO cells (clones #1 and #2), and STAT2 KO cells (clone #1) were treated with IFN-α (1,000 U/ml) or IFN-λ (1,000 U/ml) for the indicated times. The levels of PKR and MX1 mRNAs were evaluated by real-time PCR and normalized to control values. Data represent the mean ± S.D. ( n = 3). ( C ) NT sgRNA-expressing cells and STAT1 KO cells (clone #1) were treated with IFN-α (1,000 U/ml) or IFN-λ (1,000 U/ml) for 24 h. Microarray analysis was performed. Fold changes relative to untreated NT sgRNA-expressing cells were calculated. Probe sets that showed >1.5-fold increase in response to both IFN-α and IFN-λ in NT sgRNA-expressing cells but showed little changes (within 1.5-fold) due to STAT1 KO were selected. Heat maps were generated using the microarray data. See for a full list of the selected probe sets and fold changes.

Article Snippet: Anti-phospho-STAT2 (600-401-A93S) antibody was purchased from Rockland Immunochemicals.

Techniques: Expressing, Knock-Out, Infection, Western Blot, Clone Assay, Real-time Polymerase Chain Reaction, Control, Microarray, Generated

Journal: Scientific Reports

Article Title: STAT1 is essential for the inhibition of hepatitis C virus replication by interferon-λ but not by interferon-α

doi: 10.1038/srep38336

Figure Lengend Snippet: ( A , B ) NT sgRNA-expressing Huh-7.5 cells, STAT1 knockout (KO) cells (clones #1 and #2), and STAT2 KO cells (clone #1) were treated with IFN-α (1,000 U/ml) ( A ) or IFN-λ (1,000 U/ml) ( B ) for the indicated times. Phosphorylation of STAT1 and STAT2 was evaluated by immunoblotting (IB).

Article Snippet: Anti-phospho-STAT2 (600-401-A93S) antibody was purchased from Rockland Immunochemicals.

Techniques: Expressing, Knock-Out, Clone Assay, Phospho-proteomics, Western Blot

Journal: Cell reports

Article Title: Symmetric Arginine Dimethylation Is Selectively Required for mRNA Splicing and the Initiation of Type I and Type III Interferon Signaling.

doi: 10.1016/j.celrep.2020.01.054

Figure Lengend Snippet: Figure 6. Targeted Exclusion of ATF2 Exon 5 Alone Is Insufficient to Repress IFN Signaling Induction (A–C) Percentage spliced in (PSI) of ATF2e5 in naive CD4+ (A) or naive CD8+ (B) T cells activated with DMSO, 10 mM EPZ, 300 nM PF-9927, or 300 nM PF-9924 or THP-1 cells (C) treated with DMSO or 300 nM PF-9927 for 72 h. (D) Left: representative RT-PCR plot of THP-1 cells transfected with ATF2e5-targeting ASO1. Right: percentage of ATF2e5 skipping that occurs in THP-1 cells 48 h post-transfection with three ATF2e5-targeting ASOs. (E–G) Correlation of ATF2 protein expression (E), IFNB1 expression (F, top), IFNL1 expression (F, bottom), IFNb production (G, top), and IFNl1 production (G, bottom) with percentage of skipped ATF2e5 when THP-1 cells are transfected with ATF2e5-targeting ASOs for 24 h and stimulated overnight with 2030-cGAMP. All samples were normalized to non-targeting ASO controls within each experimental replicate. (H) Expression and phosphorylation of STAT1, STAT2, and IFIT1 when THP-1 cells are transfected and stimulated as in (E)–(G). Each point represents an individual ASO treatment, and bars represent mean ± SD. GAPDH or actin were used as loading controls. All data are pooled from three independent experiments, except (E) and (H), which are representative of two independent experiments with two ATF2e5-targeting ASOs. */#p < 0.05, **/##p < 0.01, ***/###p < 0.001, ****/####p < 0.0001, relative to DMSO or non-targeting controls/PF-9924 controls. (A, B, and D: one-way ANOVA with Tukey’s multiple comparison test; C: two-tailed unpaired t test). (E)–(G) show R2 and p values calculated from linear regressions.

Article Snippet: STAT2 (MAB1666, 1:1000) antibody was purchased from R&D Systems.

Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Expressing, Phospho-proteomics, Comparison, Two Tailed Test