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p irf3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p irf3
    P Irf3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+irf3/pmc12989726-434-17-26
    Average 86 stars, based on 1 article reviews
    p irf3 - by Bioz Stars, 2026-09
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    Related Articles

    Western Blot:

    Article Title: ISGylation prevents autophagic degradation of STING and promotes antitumor immunity in lung cancer
    Article Snippet: For transient transfection experiments, cells at 70% confluency were transfected with plasmid DNA constructs using Hieff Trans Liposomal Transfection Reagent (Yeasen, 40802). .. The antibodies used for western blotting included anti-Flag (Sigma-Aldrich, F7425 and Cell Signaling Technology, 14793), anti-Myc (Cell Signaling Technology, 18583), anti-HA (Cell Signaling Technology, 3724), anti-STING (Cell Signaling Technology, 13647), anti-ISG15 (Invitrogen, 7H29L24), anti-IRF3 (Cell Signaling Technology, 4302), anti-pIRF3 (Cell Signaling Technology, 79945), anti-LC3I/II (Cell Signaling Technology, 3868), anti-p62 (Cell Signaling Technology, 39749), anti-Ub (Cell Signaling Technology, 20326), anti-K48-Ub (Cell Signaling Technology, 8081), anti-GAPDH (Cell Signaling Technology, 5174), anti-rabbit IgG, HRP-linked antibody (Cell Signaling Technology, 7074) and anti-mouse IgG, HRP-linked antibody (Cell Signaling Technology, 7076). .. The antibodies used for flow cytometry assay included anti-mouse-CD45-FITC (BioLegend, 103107), anti-mouse-CD8a-APC (BioLegend, 100711) and anti-mouse-NK1.1-PE (BioLegend, 156503). diABZi (S8796) was purchased from Selleck, tanshinone IIA sulfonate sodium (54516ES20) was purchased from Yeasen, MG132 (T2154) and cycloheximide (T1225) were purchased from Topscience, chloroquine (14774) was purchased from Cell Signaling Technology.

    Article Title: ISGylation prevents autophagic degradation of STING and promotes antitumor immunity in lung cancer.
    Article Snippet: For transient transfection experiments, cells at 70% confluency were transfected with plasmid DNA constructs using Hieff Trans Liposomal Transfection Reagent (Yeasen, 40802). .. The antibodies used for western blotting included anti-Flag (Sigma-Aldrich, F7425 and Cell Signaling Technology, 14793), anti-Myc (Cell Signaling Technology, 18583), anti-HA (Cell Signaling Technology, 3724), anti-STING (Cell Signaling Technology, 13647), anti-ISG15 (Invitrogen, 7H29L24), anti-IRF3 (Cell Signaling Technology, 4302), anti-pIRF3 (Cell Signaling Technology, 79945), anti-LC3I/II (Cell Signaling Technology, 3868), anti-p62 (Cell Signaling Technology, 39749), anti-LC3I/II (Cell Signaling Technology, 3868), anti-p62 (Cell Signaling Technology, 39749), anti-Ub (Cell Signaling Technology, 20326), anti-K48-Ub (Cell Signaling Technology, 8081), anti-GAPDH (Cell Signaling Technology, 5174), anti-rabbit IgG, HRP-linked antibody (Cell Signaling Technology, 7074) and anti-mouse IgG, HRP-linked antibody (Cell Signaling Technology, 7076). .. The antibodies used for flow cytometry assay included anti-mouseCD45-FITC (BioLegend, 103107), anti-mouse-CD8a-APC (BioLegend, 100711) and anti-mouseNK1.1-PE (BioLegend, 156503). diABZi (S8796) was purchased from Selleck, tanshinone IIA sulfonate sodium (54516ES20) was AR TI CL E IN P RE SS purchased from Yeasen, mg132 (T2154) and cycloheximide (T1225) were purchased from Topscience, chloroquine (14774) was purchased from Cell Signaling Technology.

    Membrane:

    Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling
    Article Snippet: The membrane was blocked with 5% (w/v) Nonfat Dry Milk (Cell Signaling) in tris-buffered saline and 0.1% (v/v) Tween 20 (TBST) for 1 h at RT with gentle orbital shaking. .. The membrane was incubated with primary antibodies anti-STING (1:1000 dilution, Cell Signaling #13647), anti-TBK1 (1:1000, Cell Signaling #3504), anti-phospho-TBK1 (Ser172) (1:1000, Cell Signaling #5483), anti-IRF3 (1:1000, Cell Signaling #4302), or anti-phospho-IRF3 (Ser396) (1:1000, Cell Signaling #29047) in 5% (w/v) BSA in TBST overnight at 4 °C with gentle orbital shaking. .. The membrane was incubated with the secondary antibody anti-rabbit IgG, HRP (1:2000, Cell Signaling #7074) in 5% (w/v) Nonfat Dry Milk in TBST for 1 h at RT with gentle orbital shaking.

    Incubation:

    Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling
    Article Snippet: The membrane was blocked with 5% (w/v) Nonfat Dry Milk (Cell Signaling) in tris-buffered saline and 0.1% (v/v) Tween 20 (TBST) for 1 h at RT with gentle orbital shaking. .. The membrane was incubated with primary antibodies anti-STING (1:1000 dilution, Cell Signaling #13647), anti-TBK1 (1:1000, Cell Signaling #3504), anti-phospho-TBK1 (Ser172) (1:1000, Cell Signaling #5483), anti-IRF3 (1:1000, Cell Signaling #4302), or anti-phospho-IRF3 (Ser396) (1:1000, Cell Signaling #29047) in 5% (w/v) BSA in TBST overnight at 4 °C with gentle orbital shaking. .. The membrane was incubated with the secondary antibody anti-rabbit IgG, HRP (1:2000, Cell Signaling #7074) in 5% (w/v) Nonfat Dry Milk in TBST for 1 h at RT with gentle orbital shaking.

    Article Title: STING pathway contributes to Steroid-Hyporesponsive Lung Inflammation in DSS-induced colitis mice model
    Article Snippet: .. Then, membranes were incubated with primary antibodies, anti-phospho-STING (cat. #72971, 1:1000, mouse), anti-STING (cat. #50494, 1:1000), anti-phospho-TBK1/NAK (cat. #5483,1:1000), anti-TBK1/NAK (cat. #3504, 1:1000), anti-phospho-IRF3 (cat. #29047, 1:1000), anti-IRF3 (cat. #4302, 1:1000), and β actin (cat. #4970, 1:1000) (Cell Signaling Technologies, Danvers, MA), and anti-glucocorticoid receptor α (PA1–516), anti-glucocorticoid receptor β (PA3–514) (Thermo Fisher Scientific, Waltham, MA) at 4 ◦ C overnight. .. The membranes were then probed with anti-rabbit IgG, horseradish peroxidase-conjugated secondary antibody (cat. #7074S, 1:1000; Cell Signaling Technologies) for 1 h at room temperature.

    Gentle:

    Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling
    Article Snippet: The membrane was blocked with 5% (w/v) Nonfat Dry Milk (Cell Signaling) in tris-buffered saline and 0.1% (v/v) Tween 20 (TBST) for 1 h at RT with gentle orbital shaking. .. The membrane was incubated with primary antibodies anti-STING (1:1000 dilution, Cell Signaling #13647), anti-TBK1 (1:1000, Cell Signaling #3504), anti-phospho-TBK1 (Ser172) (1:1000, Cell Signaling #5483), anti-IRF3 (1:1000, Cell Signaling #4302), or anti-phospho-IRF3 (Ser396) (1:1000, Cell Signaling #29047) in 5% (w/v) BSA in TBST overnight at 4 °C with gentle orbital shaking. .. The membrane was incubated with the secondary antibody anti-rabbit IgG, HRP (1:2000, Cell Signaling #7074) in 5% (w/v) Nonfat Dry Milk in TBST for 1 h at RT with gentle orbital shaking.



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    (A) A schematic illustrates the activation of STING, TBK1, and <t>IRF3</t> in response to double-stranded DNA (dsDNA) in the cytosol by cGAS. (B-D) THP-1 macrophages were mock-infected or infected with HSV-1 in the presence or absence of 1 µM PGE 2 . At the indicated h.p.i, cell lysates were collected and subjected to immunoblotting with the indicated antibodies (B) . Band intensity of phosphorylated TBK1 (C) and IRF3 (D) was quantified and normalized to total TBK1 and IRF3, respectively (n = 3). (E) THP-1 macrophages expressing ISRE-Luciferase were transfected with viral DNA at indicated concentrations in the presence or absence of 1 µM PGE 2 for 16 hours. Cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (F) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for the indicated times. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. Samples transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) were included as controls. (G) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1). At 16 h.p.i, cell lysates were collected and subjected to qPCR to assess levels of total mt-D-loop and mt-ND1 regions (n = 3). (H) Representative Airyscan live-cell imaging of THP-1 macrophages mock-infected or infected with HSV-1 for 3 hours. MitoTracker DeepRed (magenta): mitochondria, PicoGreen (yellow): DNA. Scale bars, 2 µm. (I) Quantification of cytosolic PicoGreen in THP-1 macrophages mock-infected or infected with HSV-1 (MOI = 1) for 3 hours (n = 10). (J-K) THP-1 macrophages were infected with HSV-1 in the presence or absence of the mtDNA replication inhibitor, ddC. At 24 h.p.i, cell supernatants were collected and subjected to ELISA to measure levels of secreted IFNβ (n = 3) (J) , and cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3) (K) . (L) THP-1 macrophages expressing ISRE-Luciferase were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2. At 16 hours post stimulation, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (M) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2 . At 16 hours post-stimulation, cell lysates were collected and subjected to RT-qPCR to assess the mRNA levels of representative type I ISGs (n = 3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test ( C-E, L-M ) or unpaired, two-tailed Student’s t-test ( G, I-K ). p -values are indicated.
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    Image Search Results


    (A) A schematic illustrates the activation of STING, TBK1, and IRF3 in response to double-stranded DNA (dsDNA) in the cytosol by cGAS. (B-D) THP-1 macrophages were mock-infected or infected with HSV-1 in the presence or absence of 1 µM PGE 2 . At the indicated h.p.i, cell lysates were collected and subjected to immunoblotting with the indicated antibodies (B) . Band intensity of phosphorylated TBK1 (C) and IRF3 (D) was quantified and normalized to total TBK1 and IRF3, respectively (n = 3). (E) THP-1 macrophages expressing ISRE-Luciferase were transfected with viral DNA at indicated concentrations in the presence or absence of 1 µM PGE 2 for 16 hours. Cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (F) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for the indicated times. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. Samples transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) were included as controls. (G) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1). At 16 h.p.i, cell lysates were collected and subjected to qPCR to assess levels of total mt-D-loop and mt-ND1 regions (n = 3). (H) Representative Airyscan live-cell imaging of THP-1 macrophages mock-infected or infected with HSV-1 for 3 hours. MitoTracker DeepRed (magenta): mitochondria, PicoGreen (yellow): DNA. Scale bars, 2 µm. (I) Quantification of cytosolic PicoGreen in THP-1 macrophages mock-infected or infected with HSV-1 (MOI = 1) for 3 hours (n = 10). (J-K) THP-1 macrophages were infected with HSV-1 in the presence or absence of the mtDNA replication inhibitor, ddC. At 24 h.p.i, cell supernatants were collected and subjected to ELISA to measure levels of secreted IFNβ (n = 3) (J) , and cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3) (K) . (L) THP-1 macrophages expressing ISRE-Luciferase were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2. At 16 hours post stimulation, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (M) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2 . At 16 hours post-stimulation, cell lysates were collected and subjected to RT-qPCR to assess the mRNA levels of representative type I ISGs (n = 3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test ( C-E, L-M ) or unpaired, two-tailed Student’s t-test ( G, I-K ). p -values are indicated.

    Journal: bioRxiv

    Article Title: The COX2-PGE2-PKA Axis Suppresses Antiviral Immunity by Inhibiting mtDNA-Dependent STING Activation

    doi: 10.64898/2026.04.03.716411

    Figure Lengend Snippet: (A) A schematic illustrates the activation of STING, TBK1, and IRF3 in response to double-stranded DNA (dsDNA) in the cytosol by cGAS. (B-D) THP-1 macrophages were mock-infected or infected with HSV-1 in the presence or absence of 1 µM PGE 2 . At the indicated h.p.i, cell lysates were collected and subjected to immunoblotting with the indicated antibodies (B) . Band intensity of phosphorylated TBK1 (C) and IRF3 (D) was quantified and normalized to total TBK1 and IRF3, respectively (n = 3). (E) THP-1 macrophages expressing ISRE-Luciferase were transfected with viral DNA at indicated concentrations in the presence or absence of 1 µM PGE 2 for 16 hours. Cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (F) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for the indicated times. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. Samples transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) were included as controls. (G) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1). At 16 h.p.i, cell lysates were collected and subjected to qPCR to assess levels of total mt-D-loop and mt-ND1 regions (n = 3). (H) Representative Airyscan live-cell imaging of THP-1 macrophages mock-infected or infected with HSV-1 for 3 hours. MitoTracker DeepRed (magenta): mitochondria, PicoGreen (yellow): DNA. Scale bars, 2 µm. (I) Quantification of cytosolic PicoGreen in THP-1 macrophages mock-infected or infected with HSV-1 (MOI = 1) for 3 hours (n = 10). (J-K) THP-1 macrophages were infected with HSV-1 in the presence or absence of the mtDNA replication inhibitor, ddC. At 24 h.p.i, cell supernatants were collected and subjected to ELISA to measure levels of secreted IFNβ (n = 3) (J) , and cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3) (K) . (L) THP-1 macrophages expressing ISRE-Luciferase were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2. At 16 hours post stimulation, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (M) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2 . At 16 hours post-stimulation, cell lysates were collected and subjected to RT-qPCR to assess the mRNA levels of representative type I ISGs (n = 3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test ( C-E, L-M ) or unpaired, two-tailed Student’s t-test ( G, I-K ). p -values are indicated.

    Article Snippet: Antibodies for PKA substrates (#9624), PKA Cα (#5842), PKA RIα/β (#3927), GAPDH (#2118), STING (#13647), pTBK1 S172 (5483), TBK1 (#3504), pIRF3 S396 (#4947), IRF3 (#4302), TFAM (#8076), HSP90 (#4877), pUB S65 (#62802), COX4 (#4850), pPINK1 S228 (#89010), STOML2 (#89199), Vinculin (#13901) were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, Infection, Western Blot, Expressing, Luciferase, Transfection, Reporter Assay, Activity Assay, Live Cell Imaging, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Two Tailed Test