anti-phosphorylated stat1  (Cell Signaling Technology Inc)

Journal: Journal of Clinical and Translational Hepatology

Article Title: Protective Effect of Mesaconate on Autoimmune Hepatitis via Suppression of Inflammatory Response and Oxidative Stress

doi: 10.14218/JCTH.2025.00112

Figure Lengend Snippet: (A–D) Protein levels of the phosphorylated and total forms of JAK1, JAK2, and STAT1 in mouse liver tissue (A, B) and macrophages (C, D) detected by Western blotting. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Symbols “+” and “–” denote the presence or absence of MSA/IFN-γ in the culture medium, respectively. ConA, concanavalin A; MSA, mesaconate; IFN-γ, interferon-gamma; JAK1/2, Janus tyrosine kinase 1/2; STAT1, signal transducer and activator of transcription 1; phos-JAK1/2, phosphorylated JAK1/2; phos-STAT1, phosphorylated STAT1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Antibodies used included: anti-GAPDH (AB0037, Abways), anti-IL-1β (26048-4-AP, Proteintech), anti-apoptotic B-cell lymphoma 2 (26593-1-AP, Proteintech), anti-Bcl2-associated X protein (50599-2-Ig, Proteintech), anti-cleaved-caspase-3 (#9661, Cell Signaling Technology (CST)), anti-cleaved-poly ADP ribose polymerase (#5625, CST), anti-inducible nitric oxide synthase (18985-1-AP, Proteintech), anti-Janus kinase 1 (JAK1) (66466-1-Ig, Proteintech), anti-phosphorylated JAK1 (phos-JAK1) (#74129, CST), anti-JAK2 (#3230, CST), anti-phosphorylated JAK2 (phos-JAK2) (#3771, CST), anti-signal transducer and activator of transcription 1 (STAT1) (10144-2-AP, Proteintech), and anti-phosphorylated STAT1 (phos-STAT1) (28977-1-AP, Proteintech).

Techniques: Western Blot

Journal: Frontiers in Pharmacology

Article Title: Ruxolitinib alleviates DSS-induced acute ulcerative colitis by inhibiting STAT1 phosphorylation and reducing MDSC infiltration

doi: 10.3389/fphar.2025.1572534

Figure Lengend Snippet: Effect of ruxolitinib on JAK/STAT signaling pathway-related genes and inhibition of interferon (IFN)-γ-Induced STAT1 activation by ruxolitinib. (A) Heatmap of transcription levels (fold-changes) of the JAK/STAT signaling-related genes in CCD841 and Jurkat cells after treatment with IFN-γ and ruxolitinib. (B) Nuclear extracts prepared from CCD841 cells with or without IFN-γ activation and analyzed for STAT1 activation using electrophoretic mobility shift assay (EMSA) with the STAT1-binding consensus sequence DNA probe. Ruxolitinib at 50, 250, and 500 nM was incubated with STAT1-DNA complexes. RUX: ruxolitinib.

Article Snippet: DSS was purchased from MP Biomedicals; Ruxolitinib was purchased from Selleck; The following antibodies were used: APC-conjugated anti-CD11b and PE-conjugated anti-Gr-1 (BioLegend), STAT1 and phosphorylated STAT1 (Tyr701) (Proteintech), phosphorylated STAT1 (Ser727) (Cell Signaling Technology).

Techniques: Inhibition, Activation Assay, Electrophoretic Mobility Shift Assay, Binding Assay, Sequencing, Incubation

Journal: Frontiers in Pharmacology

Article Title: Ruxolitinib alleviates DSS-induced acute ulcerative colitis by inhibiting STAT1 phosphorylation and reducing MDSC infiltration

doi: 10.3389/fphar.2025.1572534

Figure Lengend Snippet: Inhibition of STAT1 phosphorylation by ruxolitinib in immune cells and normal intestinal epithelial cells. (A) Western blot analysis of p-STAT1Y701, p-STAT1S727, STAT1, and β-actin in Jurkat, RAW and CCD841 cells treated with 50 IU/mL IFN-γ for the indicated times. (B–D) Quantitative analysis of p-STAT1Y701, p-STAT1S727, STAT1 levels in (B) Jurkat, (C) RAW, and (D) CCD841 cells from (A) . (E) Western blot analysis of p-STAT1Y701, p-STAT1S727, STAT1, and β-actin in cells pretreated with IFN-γ (8 h) followed by Ruxolitinib (RUX, 24 h) at indicated concentrations (μM). (F–H) Quantitative analysis of p-STAT1Y701, p-STAT1S727, STAT1 levels in (F) Jurkat, (G) RAW, and (H) CCD841 cells from (E) . # P < 0.05, ## P < 0.01, ### P < 0.001 vs. 0 μM RUX (IFN-γ-treated). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, * P < 0.001 vs. the group with IFN-γ concentration at 0; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. the IFN-γ-treated group with RUX concentration at 0. RUX: ruxolitinib.

Article Snippet: DSS was purchased from MP Biomedicals; Ruxolitinib was purchased from Selleck; The following antibodies were used: APC-conjugated anti-CD11b and PE-conjugated anti-Gr-1 (BioLegend), STAT1 and phosphorylated STAT1 (Tyr701) (Proteintech), phosphorylated STAT1 (Ser727) (Cell Signaling Technology).

Techniques: Inhibition, Phospho-proteomics, Western Blot, Concentration Assay

Journal: Scientific Reports

Article Title: Comparison of anti-inflammatory and anti-angiogenic effects of JAK inhibitors in IL-6 and TNFα-stimulated fibroblast-like synoviocytes derived from patients with RA

doi: 10.1038/s41598-025-94894-2

Figure Lengend Snippet: ( a ) Western blotting was performed to analyze STAT1, pSTAT1, STAT3, pSTAT3, and β-actin. ( b , c ) Comparison of the ratios of pSTAT1 to STAT1 and pSTAT3 to STAT1. Protein expression was determined by semi-quantitative analysis of digitally captured images. All JAK inhibitors significantly suppressed pSTAT1 and pSTAT3 levels compared to those in the control. Compared to TOF, BAR, PEF, UPA, and FIL suppressed pSTAT1 and pSTAT3 levels. STAT, signal transducer and activator of transcription; JAK, Janus kinase; TOF, tofacitinib; BAR, baricitinib; PEF, peficitinib; UPA, upadacitinib; FIL, filgotinib.

Article Snippet: The membrane was blocked with 5% skimmed milk in TBST at 25 °C for 30 min, incubated with antibodies against anti-STAT1, anti-phosphorylated STAT1 (pSTAT1), anti-STAT3, and anti-pSTAT3 (Cell Signaling Technology, Danvers, MA, US) at 4 °C for 12 h, and further incubated with horseradish peroxidase-conjugated goat anti-rabbit IgG secondary antibody at 25 °C for 1 h. The proteins were subsequently visualized using ECL Plus reagent (GE Healthcare Life Sciences, Little Chalfont, UK) on a chemiluminescence analyzer (LAS-3000 mini; Fujifilm, Tokyo, Japan).

Techniques: Western Blot, Comparison, Expressing, Control

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 1. IFNα/β down-regulation and STAT1 up-regulation in response to CSFV. (A,B) The mRNA levels and protein expression of IFNα and IFNβ were analyzed in PK-15 cells infected with CSFV (MOI = 1) or transfected with 50 µg poly(I:C) by RT-qPCR (A) and WB (B) analysis. (D) The mRNA levels of STAT1 were analyzed in PK-15 cells infected with the CSFV (MOI = 1) at 0, 3, 6, 9, 12, 24, 36, and 48 hpi by RT-qPCR analysis. (E) The protein levels of E1/E2, STAT1, and P-STAT1 (Tyr 701) were analyzed by WB analysis. (C,F) The relative expression values of IFNα, IFNβ, STAT1, and P-STAT1 proteins were quantified using Image J software (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Infection, Transfection, Quantitative RT-PCR, Software, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 2. STAT1 expression was up-regulated by UV-treated CSFV in vitro. (A) PK-15 cells were inoculated with CSFV and UV-treated CSFV at 101, 102, 103, and 104 TCID50. CSFV replication was assessed using an anti-CSFV E2 Ab at 48 hpi by IF analysis. CSFV gRNA (B) and STAT1 mRNA (C) levels in PK-15 cells inoculated with CSFV and UV-treated CSFV at 12, 24, 48, and 72 hpi were determined by RT-qPCR analysis. (D) CSFV E2 and STAT1 protein levels at 12, 24, 48, and 72 hpi were measured by WB analysis. (E) The relative expression values of STAT1 protein were quantified using Image J software (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, In Vitro, Quantitative RT-PCR, Software, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 3. CSFV up-regulated STAT1 expression via Erns, E1, and E2, but not the Core protein. PK- 15 cells were infected with CSFV (MOI = 1) and separately transfected with 1.0 µg of the recombinant eukaryotic expression vectors pCMV-Core-HA (A–D), pCMV-Erns-His (E–H), pCMV-E1-Myc (I–L), and pCMV-E2-Flag (M–P). PK-15 cells incubated with 100 IU/mL of IFNα served as the positive control, while PK-15 cells transfected with an empty pCMV vector were utilized as the negative control. Samples were collected 72 h later for subsequent analysis. CSFV Core (A), Erns (E), E1 (I), E2 (M), and STAT1 (B,F,J,N) mRNA levels were measured by RT-qPCR analysis at 12, 24, 48, and 72 h. (C,G,K,O) CSFV E2, Core-HA, Erns-His, E1-Myc, E2-Flag, and STAT1 protein levels at 12, 24, 48, and 72 h were measured by WB analysis. (D,H,L,P) The relative expression values of STAT1 protein were quantified using Image J software (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, Infection, Transfection, Recombinant, Incubation, Positive Control, Plasmid Preparation, Negative Control, Quantitative RT-PCR, Software, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 4. STAT1 translocation to the nucleus induced by CSFV Erns, E1, and E2. PK-15 cells were used to express Erns, E1, and E2 proteins by transfecting with the pCMV-Erns-His, pCMV-E1-Myc, and pCMV-E2-Flag vectors, respectively. (A,C,E) The protein levels of STAT1 in the cytoplasm and nucleus of PK-15 cells were measured by WB analysis. (B,D,F) The relative expression values of STAT1 protein were quantified using Image J software. (G) HEK 293T cells were separately transfected with the pcDNA3.0-STAT1-His, pCMV-Erns-His, pCMV-E1-Myc, and pCMV-E2-Flag vectors, and treated with IFNα (100 IU/mL). The expression of STAT1 (a–f) and P-STAT1 (g–l) proteins were evaluated at 48 h using anti-STAT1 and anti-P-STAT1 antibodies by IFA analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Translocation Assay, Expressing, Software, Transfection, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 5. Identification of genes upstream of the STAT1 pathway during CSFV infection. (A) A heatmap of changes to the transcriptional levels of IL-8, IL-10, TGFβ1, TGFβ3 CCR2, CCR3, IFNγ, and STAT1 in PBMCs of piglets infected with CSFV. (B) PK-15 cells were infected with CSFV (MOI = 1) and the CSFV gRNA copy number was measured by RT-qPCR analysis. (C) The transcriptional levels of porcine IL-8, IL-10, TGFβ1, TGFβ3 CCR2, CCR3, and IFNγ at 12, 24, 48, and 60 hpi were measured by RT-qPCR analysis. (D, E) The relative protein expression of CSFV E2 (D) and IL-10 (E) were measured by ELISA (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Infection, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 7. IL-10 up-regulated STAT1 expression in vitro. (A,B) Following porcine IL-10 (100 IU/mL) and porcine IFNα (100 IU/mL) treatment, the transcription and protein levels of STAT1 at 12, 24, 36, and 48 h were measured by RT-qPCR and WB analysis. (C) The relative expression values of STAT1 protein were quantified using Image J software. (D) Subcellular localization of STAT1 in PK-15 cells separately treated with IL-10 (100 IU/mL) and IFNα (100 IU/mL). The expression of STAT1 (a–c) and P-STAT1 (d–f) proteins were evaluated at 48 h using anti-STAT1 and anti-P-STAT1 antibodies by IFA analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Expressing, In Vitro, Quantitative RT-PCR, Software, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 8. Knockdown of IL-10 down-regulated STAT1 expression. The transcription and protein levels of Erns, E1, E2, IL-10, and STAT1 have been detected by RT-qPCR, ELISA, and WB analysis in PK-15 cells temporarily transfected with IL-10 siRNA or control siRNA (100 nM) for 6 h, and then transfected with the pCMV-Erns-His (A–F), pCMV-E1-Myc (G–L), or pCMV-E2-Flag (M–R) vectors, respectively, underlying the transfection of the empty vector pCMV (1.0 µg) as a negative control at 12, 24, 36, and 48 hpt. The mRNA levels of Erns, E1, and E2 (A,G,M), IL-10 (B,H,N), and STAT1 (D,J,P) are displayed by RT-qPCR analysis. The protein levels of IL-10 (C,I,O) and STAT1 (E,K,Q) were detected by ELISA and WB analysis. (F,L,R) The relative expression values of the STAT1 protein were quantified using Image J software (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Transfection, Control, Plasmid Preparation, Negative Control, Software, Western Blot

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 9. Identification of downstream targets of the STAT1 pathway. (A,B) The copy number of CSFV gRNA and the mRNA levels of porcine MX1, OAS1, and ISG20 were quantified through RT-qPCR analysis at 12, 24, 36, and 48 hpi in PK-15 cells infected with CSFV (MOI = 1). (C) The protein levels were assessed using Western blot (WB) analysis. (D) The mRNA levels of porcine STAT1, MX1, OAS1, and ISG20 were quantified using RT-qPCR analysis at 12, 24, 36, and 48 hpt in PK-15 cells transfected with the pcDNA3.0-STAT1-His vector, and (E) protein levels were assessed through WB analysis. (F) The mRNA levels of porcine STAT1, MX1, OAS1, and ISG20 were quantified using RT-qPCR analysis at 12, 24, 36, and 48 hpi in CSFV-infected PK-15STAT1-/- cells and (G) protein levels were measured by WB analysis (** p < 0.01, *** p < 0.001, **** p < 0.0001). Original images of the western blots can be found at Supplementary Materials.

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Quantitative RT-PCR, Infection, Western Blot, Transfection, Plasmid Preparation

Journal: Biomolecules

Article Title: Classical Swine Fever Virus Envelope Glycoproteins E rns , E1, and E2 Activate IL-10-STAT1-MX1/OAS1 Antiviral Pathway via Replacing Classical IFNα/β.

doi: 10.3390/biom15020200

Figure Lengend Snippet: Figure 10. Substitution of the classical IFNα/β-STAT1 pathway with the IL-10-STAT1 pathway during CSFV infection. The type I IFN-mediated STAT1 pathway was replaced with the alternative IL-10-STAT1 pathway during CSFV infection via the viral envelope glycoproteins Erns, E1, and E2. The classical pathway: IFNα/β-p-STAT1/p-STAT2-IRF9-ISGs (left panel). The alternative pathway: IL-10-p-STAT1-MX1/OAS1 (right panel).

Article Snippet: Following blocking, the membranes were probed at 37 ◦C for 60 min with primary antibodies (Abs) against STAT1 (10144-2-AP), Myc tag (16286-1-AP), lamin B1 (12987-1-AP), OAS1 (14955-1-AP), MX1 (13750-1-AP), His tag (10001-0-AP), and Flag tag (20543-1-AP) (all from Proteintech Co., Ltd., Wuhan, China); phosphorylated (p) STAT1 (9167S), IFNα (6B18), and IFNβ (D2J1D) (from Cell Signaling Technology, Inc., Danvers, MA, USA); CSFV E1/E2 (9011) (from JENO Biotech, Seoul, Korea); CSFV E2 (orb500382) (from Biorbyt Ltd., Cambridge, England); and β-actin (CW0096A) (from Beijing CoWin Biotech Co., Ltd., Beijing, China).

Techniques: Infection