Journal: Molecular Metabolism

Article Title: Baricitinib counteracts metaflammation, thus protecting against diet-induced metabolic abnormalities in mice

doi: 10.1016/j.molmet.2020.101009

Figure Lengend Snippet: Baricitinib attenuates the HD-induced JAK-STAT pathway. Western blotting analysis for phosphorylation of Tyr 1007/1008 JAK1/2 in the skeletal muscle (A) and kidneys (B) and normalized to total JAK1/2 and for phosphorylation of Tyr 690 on STAT2 in the skeletal muscle (C) and kidneys (D) and normalized to total STAT2. All of the data are expressed as mean ± SEM for n = 6 per group. ∗p< 0.05 vs ND and •p< 0.05 vs HD.

Article Snippet: The antibodies used were rabbit anti-Tyr 1007/1008 JAK2 (#3776), rabbit anti-total JAK2 (#3230), rabbit anti-Tyr 690 STAT2 (Bioss Antibodies, bs-3428R), rabbit anti-total STAT2 (#72604), rabbit p21 (#2947), rabbit anti-Ser 307 IRS-1 (#2381), mouse anti-total IRS-1 (#3194), rabbit anti-Ser 473 AKT (#4060), rabbit anti-total AKT (#9272), rabbit anti-Ser 9 GSK-3β (#9332) and rabbit anti-total GSK-3β (9315).

Techniques: Western Blot

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 2 ASFV pA104R inhibits ISGF3-induced ISRE promoter activity and attenuates the phosphorylation of STAT1. (A) HEK-293 T cells were transfected with various concentrations of pA104R plasmids, along with ISGF3 complex (STAT1, STAT2 and IRF9) and pISRE-Luc and pRL- TK plasmids. After 30 h, a luciferase assay was performed. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. (B) HEK-293 T cells were transfected with pA104R or empty vector. After 24 h, cells were treated with 1,000 U/ ml IFN-α for 2 h. The levels of total or phosphorylated STAT1, STAT2, and IRF9 were detected by immunoblotting analysis.

Article Snippet: The STAT1 (9172), STAT2 (72604), phosphor-STAT1 (9649), phosphor-STAT2 (88410), and IRF9 (76684) antibodies were purchased from Cell Signaling Technology (Danvers, MA). β-Actin (66009-1-Ig), GFP-tag (66002-1-Ig), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP) antibodies were purchased from Proteintech (Chicago, IL).

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

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 3 ASFV pA104R does not interact with STAT1, STAT2, and IRF9. HEK-293 T cells were transfected with pA104R alone (C) or co-transfected with STAT1, STAT2, and IRF9 (A,B). After 24 h, cells were treated with 1,000 U/mL IFN-α for 8 h. Cell lysates were prepared and subjected to immunoprecipitation analysis. The whole-cell lysates and immunoprecipitation complexes were analyzed by immunoblotting with the indicated antibodies.

Article Snippet: The STAT1 (9172), STAT2 (72604), phosphor-STAT1 (9649), phosphor-STAT2 (88410), and IRF9 (76684) antibodies were purchased from Cell Signaling Technology (Danvers, MA). β-Actin (66009-1-Ig), GFP-tag (66002-1-Ig), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP) antibodies were purchased from Proteintech (Chicago, IL).

Techniques: Transfection, Immunoprecipitation, Western Blot

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 5 ASFV pA104R does not prevent the association of ISGF3 with promoter. (A) The ISRE DNA and control oligonucleotides used for DNA pull-down. (B) HEK-293T cells were co-transfected with ISGF3 complex (STAT1, STAT2, and IRF9), along with pA104R or empty control. After 24 h post- transfection, cells were treated with 1,000 U/mL IFN-α for 12 h. Nuclear extracts were incubated with a Biotin-labeled ISRE or control probe and subjected to pull-down analysis with streptavidin magnetic beads. The whole-cell lysates and pull-down complexes were analyzed by immunoblotting with the indicated antibodies.

Article Snippet: The STAT1 (9172), STAT2 (72604), phosphor-STAT1 (9649), phosphor-STAT2 (88410), and IRF9 (76684) antibodies were purchased from Cell Signaling Technology (Danvers, MA). β-Actin (66009-1-Ig), GFP-tag (66002-1-Ig), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP) antibodies were purchased from Proteintech (Chicago, IL).

Techniques: Control, Transfection, Incubation, Labeling, Magnetic Beads, Western Blot

Journal: Frontiers in microbiology

Article Title: African swine fever virus pA104R protein acts as a suppressor of type I interferon signaling.

doi: 10.3389/fmicb.2023.1169699

Figure Lengend Snippet: FIGURE 6 ASFV pA104R does not exert repressive effects on trans-activation domains and transcriptional co-stimulatory factors. (A) HEK-293 T cells were co- transfected with pA104R and IRF9-Stat2(TA) (A) or GAL4-Stat2(TA) (B), along with pRL-TK and pISRE-Luc (A) or pGAL4-UAS-Luc (B) plasmids. After 30 h, a luciferase assay was performed. (C) HEK-293 T Cells were co-transfected with pA104R and CBP or p300 along with pISRE-Luc and pRL-TK plasmids. After 24 h post-transfection, cells were treated with 1,000 U/mL IFN-α for 12 h, followed by luciferase assays. Data are shown as means ± SEM from three independent experiments. Statistical analysis was performed by one-way ANOVA. *** p < 0.001.

Article Snippet: The STAT1 (9172), STAT2 (72604), phosphor-STAT1 (9649), phosphor-STAT2 (88410), and IRF9 (76684) antibodies were purchased from Cell Signaling Technology (Danvers, MA). β-Actin (66009-1-Ig), GFP-tag (66002-1-Ig), Flag-tag (20543-1-AP), and HA-tag (51064-2-AP) antibodies were purchased from Proteintech (Chicago, IL).

Techniques: Activation Assay, Transfection, Luciferase

Journal: Heliyon

Article Title: Innate antiviral responses against Shaan virus infection in HEK293, A549 and MARC-145 cells and limited role of viperin against Shaan virus replication

doi: 10.1016/j.heliyon.2023.e22597

Figure Lengend Snippet: The expression level of IFN-α, -β and -λ1 in Shaan virus-infected HEK293, A549 and MARC-145 cells HEK293, A549, and MARC-145 cells were infected with the Shaan virus (ShaV) at an MOI of 1 or mock. RNA expression levels of IFN-α (A), -β (B) and -λ1 (C) were detected using real time RT-PCR. Data were normalized to GAPDH expression and presented as relative fold expression values for each control cell. P values of <0.05 were considered statistically significant and denoted as p < 0.05 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****) (n = 3). (D) Western blots of STAT1 and STAT2 proteins in infected cells at the indicated time points. Uncropped images of western blots displayed in .

Article Snippet: The following primary antibodies were used in this study: IRF3 (D83B9), IRF7 (D2A1J), phosphorylated-IRF3 Ser396 (4D4G), STAT1 (D1K9Y), STAT2 (D9J7L), phosphorylated- STAT1 Tyr701 (58D6) and phosphorylated- STAT2 Tyr690 (D3P2P) (Cell signaling Technology), viperin (ab107359), and beta-actin (ab8226) (Abcam).

Techniques: Expressing, Virus, Infection, RNA Expression, Quantitative RT-PCR, Control, Western Blot

Journal: Heliyon

Article Title: Innate antiviral responses against Shaan virus infection in HEK293, A549 and MARC-145 cells and limited role of viperin against Shaan virus replication

doi: 10.1016/j.heliyon.2023.e22597

Figure Lengend Snippet: IRF3 phosphorylation and IRF7 induction in Shaan virus-infected cells HEK293, A549, and MARC-145 cells were infected with the Shaan virus at an MOI of 1. Lysates from uninfected MARC-145 cells were used as controls. The IRF3 protein (A) and IRF7 protein (B) were detected by western blotting at the indicated time points. Uncropped images of western blots displayed in .

Article Snippet: The following primary antibodies were used in this study: IRF3 (D83B9), IRF7 (D2A1J), phosphorylated-IRF3 Ser396 (4D4G), STAT1 (D1K9Y), STAT2 (D9J7L), phosphorylated- STAT1 Tyr701 (58D6) and phosphorylated- STAT2 Tyr690 (D3P2P) (Cell signaling Technology), viperin (ab107359), and beta-actin (ab8226) (Abcam).

Techniques: Phospho-proteomics, Virus, Infection, Western Blot