rabbit α p stat1 y701  (Cell Signaling Technology Inc)

Journal: Cell Insight

Article Title: HTATSF1 regulates innate antiviral immune response by orchestrating TRAF3-IRF3 and TRAF6-NF-κB pathways

doi: 10.1016/j.cellin.2025.100294

Figure Lengend Snippet: HTATSF1 mediates viral nucleic acid- but not IFN-β-triggered signaling. (A) Effects of HTATSF1-deficiency on transcription of downstream genes induced by viral nucleic acid mimics. The control (g NC ) and HTATSF1-deficient (g HTATSF1 ) THP-1 cells (1 × 10 6 ) were untransfected or transfected with HT-DNA (2 μg/mL) or poly(I:C) (2 μg/mL) for the indicated times before RT-qPCR analysis to measure mRNA levels of the indicated genes. (B) Effects of HTATSF1-deficiency on transcription of downstream genes induced by cGAMP. The control (g NC ) and HTATSF1-deficient (g HTATSF1 ) THP-1 cells (1 × 10 6 ) were left untreated or treated with 2′3′-cGAMP (40 nM) for the indicated times before RT-qPCR analysis to measure mRNA levels of the indicated genes. (C) Effects of HTATSF1-deficiency on IFN-β-induced phosphorylation of STAT1. The control (g NC ) and HTATSF1-deficient (g HTATSF1 ) THP-1 cells (1 × 10 6 ) were left untreated or treated with IFN-β (100 ng/mL) for the indicated times before immunoblotting analysis with the indicated antibodies. (D) Effects of HTATSF1-deficiency on transcription of downstream genes induced by IFN-β. The control (g NC ) and HTATSF1-deficient (g HTATSF1 ) THP-1 cells (1 × 10 6 ) were left untreated or treated with IFN-β (100 ng/mL) for 2 h before RT-qPCR analysis to measure mRNA levels of the indicated genes. Data shown are mean ± SEM ( n = 3 technical replicates) from one representative experiment ( (A) , ( B) , (D) ), which were repeated for at least two times with similar results. n.s., not significant; ∗∗, P < 0.01 (unpaired t -test).

Article Snippet: Antibodies against HA (TA180128) (OriGene); Flag (F3165) and β-actin (A2228) (Sigma); p-TBK1 S172 (ab109272), TBK1 (ab40676), p-IRF3 S386 (ab76493), TAK1 (ab109526), TRAF6 (ab33915), p65 (ab7970), ubiquitin (ab7254), K48-linkage specific polyubiquitin (ab140601) and K63-linkage specific polyubiquitin (ab179434) (Abcam); IRF3 (sc-33641) and STAT1 (SC-417) (Santa Cruz Biotechnology); Myc (5605), p-IRF3 S396 (4947), p-STAT1 Y701 (9167), TRAF3 (4729), p-TAK1 T184/187 (4508), p-IκBα S32/36 (9246), IκBα (9242), p-IKKα/β S176/177 (2078), IKKβ (2370), and p-p65 S536 (3033) (Cell Signaling Technology), HTATSF1 (20805-1-AP) and HECTD3 (11487-1-AP) (Proteintech) were purchased from the indicated companies.

Techniques: Control, Transfection, Quantitative RT-PCR, Phospho-proteomics, Western Blot

Journal: Cell Insight

Article Title: HTATSF1 regulates innate antiviral immune response by orchestrating TRAF3-IRF3 and TRAF6-NF-κB pathways

doi: 10.1016/j.cellin.2025.100294

Figure Lengend Snippet: HTATSF1 is important for HSV-1- or SeV-triggered phosphorylation of downstream signaling components. (A) Effects of HTATSF1-deficiency on HSV-1- or SeV-induced phosphorylation of TBK1, IRF3, and STAT1 in THP-1 cells. The control (g NC ) and HTATSF1-deficient (g HTATSF1 ) THP-1 cells (1 × 10 6 ) were left uninfected or infected with HSV-1 (MOI = 1) or SeV (MOI = 1) for the indicated times before immunoblotting analysis with the indicated antibodies. (B) Effects of HTATSF1-deficiency on HSV-1- or SeV-induced phosphorylation of TBK1, IRF3, and STAT1 in BMDMs. Lyz2 -Cre; Htatsf1 fl/fl and Htatsf1 fl/fl BMDM cells (1 × 10 6 ) were left uninfected or infected with HSV-1 (MOI = 1) or SeV (MOI = 1) for the indicated times before immunoblotting analysis with the indicated antibodies.

Article Snippet: Antibodies against HA (TA180128) (OriGene); Flag (F3165) and β-actin (A2228) (Sigma); p-TBK1 S172 (ab109272), TBK1 (ab40676), p-IRF3 S386 (ab76493), TAK1 (ab109526), TRAF6 (ab33915), p65 (ab7970), ubiquitin (ab7254), K48-linkage specific polyubiquitin (ab140601) and K63-linkage specific polyubiquitin (ab179434) (Abcam); IRF3 (sc-33641) and STAT1 (SC-417) (Santa Cruz Biotechnology); Myc (5605), p-IRF3 S396 (4947), p-STAT1 Y701 (9167), TRAF3 (4729), p-TAK1 T184/187 (4508), p-IκBα S32/36 (9246), IκBα (9242), p-IKKα/β S176/177 (2078), IKKβ (2370), and p-p65 S536 (3033) (Cell Signaling Technology), HTATSF1 (20805-1-AP) and HECTD3 (11487-1-AP) (Proteintech) were purchased from the indicated companies.

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

Journal: Scientific reports

Article Title: Caerin 1.1/1.9-mediated antitumor immunity depends on IFNAR-Stat1 signalling of tumour infiltrating macrophage by autocrine IFNα and is enhanced by CD47 blockade.

doi: 10.1038/s41598-025-87687-0

Figure Lengend Snippet: Fig. 4. The anti-tumor effect of F1/F3 relies on the STAT1 related axis. (A) Experimental timeline and design for F1/F3 treatment. (B) Levels of p-STAT1 (Y701) in DCs (CD45.2+CD11c+MHCII+) and macrophages (CD45.2+CD11b+F4/80+) were assessed following F1/F3, control P3 peptide, or saline treatment, using intracellular staining (ICS) and flow cytometry. (C) Survival rates and tumour volumes in treated and control mice administered 2 mg fludarabine or 100 µl PBS intraperitoneally every other day. (E) Survival analysis of mice with STAT1 inhibition in conjunction with F1/F3 treatment. Each experiment included 4–10 mice per group, with data representative of one independent experiment, shown as mean ± SD or mean ± SEM (D). *P < 0.05; **P < 0.01; ***P < 0.001; by one-way ANOVA test (B) or simple survival analysis (Kaplan–Meier) (C,E) or two-way ANOVA test (D).

Article Snippet: Antibodies against p-Stat1 (Y701) (clone: D4A7), p-Stat1 (S727) (clone: 8826), β-actin were used in this study, all of which were purchased from Cell Signalling Technology.

Techniques: Control, Saline, Staining, Flow Cytometry, Inhibition

Journal: Scientific reports

Article Title: Caerin 1.1/1.9-mediated antitumor immunity depends on IFNAR-Stat1 signalling of tumour infiltrating macrophage by autocrine IFNα and is enhanced by CD47 blockade.

doi: 10.1038/s41598-025-87687-0

Figure Lengend Snippet: Fig. 7. Schematic diagram of mechanisms activated by caerin peptide therapy. Following Caerin peptide administration, there is an increase in immune cell infiltration into the tumour, including T cells, macrophages, and CD45.2+ cells. Activated macrophages secrete IFNα, which initiates the IFNAR-1/STAT1 signalling pathway. This pathway boosts macrophage sensitivity to type I IFN, enhancing their function. Consequently, macrophages polarise into an inflammatory phenotype with potent anti-tumour activity, contributing to effective tumour treatment.

Article Snippet: Antibodies against p-Stat1 (Y701) (clone: D4A7), p-Stat1 (S727) (clone: 8826), β-actin were used in this study, all of which were purchased from Cell Signalling Technology.

Techniques: Activity Assay

Journal: Scientific Reports

Article Title: Caerin 1.1/1.9-mediated antitumor immunity depends on IFNAR-Stat1 signalling of tumour infiltrating macrophage by autocrine IFNα and is enhanced by CD47 blockade

doi: 10.1038/s41598-025-87687-0

Figure Lengend Snippet: In vivo antibodies and flow cytometry antibodies.

Article Snippet: P-stat1 (Y701) , PE , lot6 , Stat1Y701-3E6 , Thermo Fisher Scientific.

Techniques: In Vivo, Flow Cytometry, Staining, Cell Stimulation, Control

Journal: Scientific Reports

Article Title: Caerin 1.1/1.9-mediated antitumor immunity depends on IFNAR-Stat1 signalling of tumour infiltrating macrophage by autocrine IFNα and is enhanced by CD47 blockade

doi: 10.1038/s41598-025-87687-0

Figure Lengend Snippet: The anti-tumor effect of F1/F3 relies on the STAT1 related axis. ( A ) Experimental timeline and design for F1/F3 treatment. ( B ) Levels of p-STAT1 (Y701) in DCs (CD45.2 + CD11c + MHCII + ) and macrophages (CD45.2 + CD11b + F4/80 + ) were assessed following F1/F3, control P3 peptide, or saline treatment, using intracellular staining (ICS) and flow cytometry. ( C ) Survival rates and tumour volumes in treated and control mice administered 2 mg fludarabine or 100 µl PBS intraperitoneally every other day. ( E ) Survival analysis of mice with STAT1 inhibition in conjunction with F1/F3 treatment. Each experiment included 4–10 mice per group, with data representative of one independent experiment, shown as mean ± SD or mean ± SEM ( D ). * P < 0.05; ** P < 0.01; *** P < 0.001; by one-way ANOVA test ( B ) or simple survival analysis (Kaplan–Meier) ( C , E ) or two-way ANOVA test ( D ).

Article Snippet: P-stat1 (Y701) , PE , lot6 , Stat1Y701-3E6 , Thermo Fisher Scientific.

Techniques: Control, Saline, Staining, Flow Cytometry, Inhibition

Journal: Scientific Reports

Article Title: Caerin 1.1/1.9-mediated antitumor immunity depends on IFNAR-Stat1 signalling of tumour infiltrating macrophage by autocrine IFNα and is enhanced by CD47 blockade

doi: 10.1038/s41598-025-87687-0

Figure Lengend Snippet: Schematic diagram of mechanisms activated by caerin peptide therapy. Following Caerin peptide administration, there is an increase in immune cell infiltration into the tumour, including T cells, macrophages, and CD45.2 + cells. Activated macrophages secrete IFNα, which initiates the IFNAR-1/STAT1 signalling pathway. This pathway boosts macrophage sensitivity to type I IFN, enhancing their function. Consequently, macrophages polarise into an inflammatory phenotype with potent anti-tumour activity, contributing to effective tumour treatment.

Article Snippet: P-stat1 (Y701) , PE , lot6 , Stat1Y701-3E6 , Thermo Fisher Scientific.

Techniques: Activity Assay