anti p irf3 antibody  (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 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: The Journal of Headache and Pain

Article Title: Spreading depolarization activates the cGAS–STING pathway and drives cranial nociception: therapeutic potential of STING modulation

doi: 10.1186/s10194-026-02267-5

Figure Lengend Snippet: Expression of cGAS-STING pathway proteins in the ipsilateral parietal cortex increases following SD. (A) Schematic representation of the experimental design. (B) Representative immunofluorescence images showing STING, p-IRF3, and IFN-β expression in the ipsilateral parietal cortex of mice which underwent sham surgery, a single or six SDs. Scale bars: 50 μm. (C) Representative lane view images of capillary-based nanoimmunoassay (WES) analysis of STING, p-STING, IRF3, p-IRF3, and IFN-β protein levels in the ipsilateral parietal cortex following SD ( n = 4/group). D-H. WES analysis of STING, p-STING, IRF3, p-IRF3, and IFN-β protein levels in the ipsilateral parietal cortex following SD ( n = 4/group). Protein levels measured by WES were normalized to vinculin, a commonly used housekeeping protein, and the data are presented as fold changes relative to the sham group. Values are mean ± SEM. Statistical comparisons were made using Student’s t test with Welch’s correction. Not significant ( ns): p > 0.05, *: p < 0.05, **: p < 0.01, ***: p < 0.001. Panel A was created using BioRender.com

Article Snippet: The sections were washed with TRIS buffered saline (TBS) and blocked with 10% normal goat serum (Jackson ImmunoResearch, 005-000-121) at room temperature for 1 h. Next, sections were incubated overnight at +4 °C with primary antibodies: rabbit anti-STING antibody (1:200, monoclonal D2P2F, 13647 S, Cell Signaling Technology), rabbit anti-cGAS antibody (1:200, polyclonal, MBS9143559, MyBioSource), rabbit anti-p-IRF3 antibody (1:300, monoclonal, 29047 S, Cell Signaling Technology), rabbit anti-IFN-β antibody (1:300, polyclonal, NBP1-77288, Novus), rabbit anti-NLRP3 antibody (1:300, polyclonal, PA5115660, Thermo Fisher Scientific), rabbit anti-p65 antibody (1:200, monoclonal, 8242 S, Cell Signaling Technology), mouse anti-NeuN antibody (1:300, monoclonal MAB377 clone A60, Merck Millipore), goat anti-Iba1 antibody (1:250, polyclonal, NB100-1028, Novus), mouse anti-S100β antibody (1:200, polyclonal, Atlas Antibodies), rabbit anti-golgi autoantigen, golgin subfamily a, 2 (GOLGA2) antibody (1:500, polyclonal, 11308-1-AP, Proteintech).

Techniques: Expressing, Immunofluorescence