phosphorylated stat1 9167s antibody  (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