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Santa Cruz Biotechnology irf 3
Irf 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/irf 3/product/Santa Cruz Biotechnology
Average 97 stars, based on 4 article reviews
Price from $9.99 to $1999.99
irf 3 - by Bioz Stars, 2020-01
97/100 stars

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Related Articles

Nucleic Acid Electrophoresis:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: Thirty micrograms of cytoplasmic protein extracts or 20 μ g of nuclear protein extracts were denatured in Laemmli buffer and separated using 8% SDS—polyacrylamide gel electrophoresis. .. After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA). .. IKKɛ and TBK1 proteins were detected using monoclonal anti-FLAG antibodies (M2; Sigma-Aldrich).

Transfection:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Additional experiments with a construct coding for IRF-3 short hairpin RNA or IRF-7 short hairpin RNA (sh-TRCN 5919 and sh-TRCN 14859, respectively; Sigma-Aldrich, MO) provided inhibitory results similar to those of a control plasmid (shc002).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: Cells were harvested 72 h after transfection and analyzed for IFN-A gene expression by reverse transcription-qPCR. .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA).

shRNA:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Additional experiments with a construct coding for IRF-3 short hairpin RNA or IRF-7 short hairpin RNA (sh-TRCN 5919 and sh-TRCN 14859, respectively; Sigma-Aldrich, MO) provided inhibitory results similar to those of a control plasmid (shc002).

Binding Assay:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. The C module is critical for IRF-3 and CBP recruitment to the mouse IFN-A4 promoter but is dispensable for IRF-7 binding, which is mediated predominantly via the B and D modules ( ). .. Sequence comparison indicated that the B module is highly conserved in human IFN-A gene promoters (Fig. ).

Methylation:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. To assess this distinct effect of IRF-3 on IFN-A genes, 2fTGH cells, a human fibrosarcoma cell line, were used ( , ), because these cells constitutively express IRF-3 and TBK1 (150 to 200 and 250 to 300 copies, respectively), but do not express IRF-7 due to high methylation of the IRF-7 gene promoter ( , ). .. Infection of 2fTGH cells by Sendai virus induced high levels of IFN-B (1,000 copies) and very low levels of IFN-A1 (2 to 4 copies) at 4 h postinfection, whereas other IFN-A mRNAs were undetected (Fig. ).

Transferring:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: .. After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA). .. Then it was incubated with secondary antibody (Amersham, Buckinghamshire, UK) and detected by ECL (Amersham).

Incubation:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA). .. Then it was incubated with secondary antibody (Amersham, Buckinghamshire, UK) and detected by ECL (Amersham).

Infection:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. A concomitant analysis of IRF-3 and IRF-7 expression revealed that IRF-3 levels (200 to 400 copies relative to 1,000 GAPDH copies) remained constant during virus infection, except for a 10-fold increase at 2 h, whereas IRF-7 levels increased dramatically after infection, reaching maximal levels at 4 h postinfection (2,500 copies); thus, the ratio of IRF-7 to IRF-3 in Namalwa cells increased from 1:1 at 2 h to 6:1 between 4 to 8 h and subsequently decreased to 2:1 (Fig. ). .. Accordingly, the IRF-7 protein was detectable at 3 to 6 h after infection and then decreased significantly thereafter, whereas IRF-3 protein levels remained essentially constant during virus infection (Fig. ).

Small Interfering RNA:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Additional experiments with a construct coding for IRF-3 short hairpin RNA or IRF-7 short hairpin RNA (sh-TRCN 5919 and sh-TRCN 14859, respectively; Sigma-Aldrich, MO) provided inhibitory results similar to those of a control plasmid (shc002).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA).

Construct:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Additional experiments with a construct coding for IRF-3 short hairpin RNA or IRF-7 short hairpin RNA (sh-TRCN 5919 and sh-TRCN 14859, respectively; Sigma-Aldrich, MO) provided inhibitory results similar to those of a control plasmid (shc002).

Expressing:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. We therefore focused on the impact of predominant IRF-3 or IRF-7 expression on IFN-A gene regulation. .. In order to modulate the levels of IRF-3 and IRF-7, it was necessary to examine IFN-A gene expression in HEK293-TLR3 cells of high transfection efficiency.

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. These results demonstrate that exclusive IFN-A1 and -B gene expression occurred when IRF-3 was stimulated by TBK-1 or IKKɛ and also indicated that overexpression of IRF-3 exerted an inhibitory effect on IFN-A gene expression. .. Furthermore, activation of multiple IFN-A genes (other than IFN-A1) was mediated predominantly through IRF-7.

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. A concomitant analysis of IRF-3 and IRF-7 expression revealed that IRF-3 levels (200 to 400 copies relative to 1,000 GAPDH copies) remained constant during virus infection, except for a 10-fold increase at 2 h, whereas IRF-7 levels increased dramatically after infection, reaching maximal levels at 4 h postinfection (2,500 copies); thus, the ratio of IRF-7 to IRF-3 in Namalwa cells increased from 1:1 at 2 h to 6:1 between 4 to 8 h and subsequently decreased to 2:1 (Fig. ). .. Accordingly, the IRF-7 protein was detectable at 3 to 6 h after infection and then decreased significantly thereafter, whereas IRF-3 protein levels remained essentially constant during virus infection (Fig. ).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. Taken together, these findings demonstrate that the differential expression of human IFN-A genes was not only due to IRF-3 and IRF-7 acting as transcriptional activators but also to an apparent inhibitory effect of IRF-3 on IRF-7-mediated transcription of IFN-A genes. .. The present study demonstrates that the modular organization of IRF-binding elements in IFN-A gene promoters dictates the differential expression of human IFN-A genes induced by virus infection.

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Cells were harvested 72 h after transfection and analyzed for IFN-A gene expression by reverse transcription-qPCR.

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. These results demonstrate that modulation of the relative ratios of IRF-3 and IRF-7 led to differential expression of IFN-A genes; surprisingly, when IRF-3 expression was predominant, IFN-A genes were poorly induced, suggesting that IRF-3 had no significant stimulatory effect or even had an inhibitory effect on IFN-A gene expression. .. Expression of increasing amounts of IRF-3 or IRF-7 in HEK293 cells further substantiated this observation (see Fig. S1 in the supplemental material).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA). .. IKKɛ and TBK1 proteins were detected using monoclonal anti-FLAG antibodies (M2; Sigma-Aldrich).

Protein Concentration:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: Protein concentration was determined using BCA reagents (Pierce) according to the manufacturer's instructions. .. After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA).

Western Blot:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: Paragraph title: 2.5. Preparation of Nuclear and Cytoplasmic Extracts for Western Blot Analysis ... After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA).

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. Specificity and efficiency of siRNA on endogenous protein expression were tested by Western blotting ( ) of whole-cell extracts (20 μg of total proteins separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis) using IRF-3 or IRF-7 polyclonal antibodies (sc-9082 and sc-9083, respectively; Santa Cruz Biotechnology, CA) and antiactin antibodies (MAB1501; Millipore Corporation, MA). .. IKKɛ and TBK1 proteins were detected using monoclonal anti-FLAG antibodies (M2; Sigma-Aldrich).

Over Expression:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: .. These results demonstrate that exclusive IFN-A1 and -B gene expression occurred when IRF-3 was stimulated by TBK-1 or IKKɛ and also indicated that overexpression of IRF-3 exerted an inhibitory effect on IFN-A gene expression. .. Furthermore, activation of multiple IFN-A genes (other than IFN-A1) was mediated predominantly through IRF-7.

BIA-KA:

Article Title: 8-Prenylkaempferol Suppresses Influenza A Virus-Induced RANTES Production in A549 Cells via Blocking PI3K-Mediated Transcriptional Activation of NF-κB and IRF3
Article Snippet: Protein concentration was determined using BCA reagents (Pierce) according to the manufacturer's instructions. .. After transferring, membrane was detected by antibodies against NF-κ B p65; Iκ B-α , IRF-3 (all purchased from Santa Cruz Biotechnology, CA, USA), Akt and c-Jun (both purchased from Cell Signaling Technology, Beverly, MA, USA).

Plasmid Preparation:

Article Title: Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿Differential Regulation of Human Interferon A Gene Expression by Interferon Regulatory Factors 3 and 7 ▿ §
Article Snippet: After 18 to 24 h, 75 ng of pcDNA3-Flag-IKKɛ or pcDNA3-Flag-TBK1 was transfected by Lipofectamine, together with 80 pmol of a small interfering RNA (siRNA) mixture that contains a pool of three target-specific siRNAs for IRF-3 (sc-35710; Santa Cruz Biotechnology, CA), IRF-7 (sc-38011), or a scrambled siRNA (sc-37007) used as a control. .. Additional experiments with a construct coding for IRF-3 short hairpin RNA or IRF-7 short hairpin RNA (sh-TRCN 5919 and sh-TRCN 14859, respectively; Sigma-Aldrich, MO) provided inhibitory results similar to those of a control plasmid (shc002).

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  • 82
    Santa Cruz Biotechnology mouse anti irf3
    ZIKV NS5 inhibits IFN-λ1-promoter activation on both <t>IRF3</t> and NF-ĸB pathways. HEK293 cells were co-transfected with expression plasmids for ∆RIG-I and ZIKV NS3 or NS5 plasmids, and with IFN-λ1-promoter-luciferase reporter plasmids which were mutated on ( A ) IRF3 or ( B ) NF-ĸB1 binding sites. An expression plasmid for HCV NS3/4A was used as a control. Luc activity was measured at 24 h after transfection. ** indicate a significant reduction in promoter activation ( p
    Mouse Anti Irf3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti irf3/product/Santa Cruz Biotechnology
    Average 82 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti irf3 - by Bioz Stars, 2020-01
    82/100 stars
      Buy from Supplier

    82
    Santa Cruz Biotechnology mouse monoclonal anti human irf3
    Neutralization of TLR2 blocks gp120-mediated IFNβ production and <t>IRF3</t> activation. ( a ) Endometrial GECs were pretreated with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibodies (all at 10 μg/ml) before exposure to gp120 (100 ng/ml) or mock treatment (media). FimH and Pam3CSK4 were used as positive controls for activation of TLR4 and TLR2, respectively. Supernatants were collected after 24 h and analyzed by ELISA for IFNβ production. Data shown are mean + s.d. and representative of three separate experiments done on cells isolated from three different tissues. ( b ) Epithelial monolayers were fixed after 2 h of exposure of gp120 with and without pretreatment with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibody and stained for IRF3. Propidium iodide was used to stain nuclei. Images were captures by a laser-scanning confocal microscopy. Magnification × 1260. Images are representative of one of three separate experiments done on cells isolated from three different tissues. ( c ) Quantitation of IRF3 colocalization were done by Image J software and presented in the graph. Significance was calculated by one-way ANOVA and IRF3 colocalization in all treatments were compared with mock treatment. * P
    Mouse Monoclonal Anti Human Irf3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti human irf3/product/Santa Cruz Biotechnology
    Average 82 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti human irf3 - by Bioz Stars, 2020-01
    82/100 stars
      Buy from Supplier

    99
    Santa Cruz Biotechnology anti irf3
    R6 competes with <t>IRF3</t> for binding to CBP. (A) Telomerized RFs were transfected with pcDNA3.1-R6HA (5, 10, or 20 μg DNA) or empty pcDNA3.1 for 40 h and subsequently transfected with poly(I · C) for 6 h. Nuclear lysates were immunoprecipitated
    Anti Irf3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti irf3/product/Santa Cruz Biotechnology
    Average 99 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    anti irf3 - by Bioz Stars, 2020-01
    99/100 stars
      Buy from Supplier

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    ZIKV NS5 inhibits IFN-λ1-promoter activation on both IRF3 and NF-ĸB pathways. HEK293 cells were co-transfected with expression plasmids for ∆RIG-I and ZIKV NS3 or NS5 plasmids, and with IFN-λ1-promoter-luciferase reporter plasmids which were mutated on ( A ) IRF3 or ( B ) NF-ĸB1 binding sites. An expression plasmid for HCV NS3/4A was used as a control. Luc activity was measured at 24 h after transfection. ** indicate a significant reduction in promoter activation ( p

    Journal: Viruses

    Article Title: Zika Virus Non-Structural Protein NS5 Inhibits the RIG-I Pathway and Interferon Lambda 1 Promoter Activation by Targeting IKK Epsilon

    doi: 10.3390/v11111024

    Figure Lengend Snippet: ZIKV NS5 inhibits IFN-λ1-promoter activation on both IRF3 and NF-ĸB pathways. HEK293 cells were co-transfected with expression plasmids for ∆RIG-I and ZIKV NS3 or NS5 plasmids, and with IFN-λ1-promoter-luciferase reporter plasmids which were mutated on ( A ) IRF3 or ( B ) NF-ĸB1 binding sites. An expression plasmid for HCV NS3/4A was used as a control. Luc activity was measured at 24 h after transfection. ** indicate a significant reduction in promoter activation ( p

    Article Snippet: Antibodies Rabbit anti-IRF3 (FL-425; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-IRF3 (SL-12; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-phospho-IRF3 (Ser396) (4D4G; Cell Signaling Technology, Danvers, MA, USA), mouse anti-HA1.1 Epitope Tag (BioLegend, San Diego, CA, USA), mouse anti-GAPDH (6C5; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-FLAG (M2; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-TBK-1/NAK (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-TBK-1/NAK (Ser172) (D52C2; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-IKKi/IKKε (Abcam, Cambridge, UK), and rabbit anti-phospho-IKKε (Ser172) (D1B7; Cell Signaling Technology, Danvers, MA, USA) were used according to the manufacturers’ instructions.

    Techniques: Activation Assay, Transfection, Expressing, Luciferase, Binding Assay, Plasmid Preparation, Activity Assay

    ZIKV NS5 inhibits IFN-λ1-promoter activation on all the components of the RIG-I pathway. HEK293 cells were co-transfected with expression plasmids for ( A ) ∆RIG-I, ( B ) MAVS, ( C ) TBK-1, ( D ) the constitutively active form of IRF3 (IRF3-5D), and ( E , F ) IKKε, and with expression plasmids for ZIKV NS3 and NS5. An expression plasmid for HCV NS3/4A was used as a control. Luc activity was measured at 24 h after transfection. ** indicate a significant reduction in promoter activation ( p

    Journal: Viruses

    Article Title: Zika Virus Non-Structural Protein NS5 Inhibits the RIG-I Pathway and Interferon Lambda 1 Promoter Activation by Targeting IKK Epsilon

    doi: 10.3390/v11111024

    Figure Lengend Snippet: ZIKV NS5 inhibits IFN-λ1-promoter activation on all the components of the RIG-I pathway. HEK293 cells were co-transfected with expression plasmids for ( A ) ∆RIG-I, ( B ) MAVS, ( C ) TBK-1, ( D ) the constitutively active form of IRF3 (IRF3-5D), and ( E , F ) IKKε, and with expression plasmids for ZIKV NS3 and NS5. An expression plasmid for HCV NS3/4A was used as a control. Luc activity was measured at 24 h after transfection. ** indicate a significant reduction in promoter activation ( p

    Article Snippet: Antibodies Rabbit anti-IRF3 (FL-425; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-IRF3 (SL-12; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-phospho-IRF3 (Ser396) (4D4G; Cell Signaling Technology, Danvers, MA, USA), mouse anti-HA1.1 Epitope Tag (BioLegend, San Diego, CA, USA), mouse anti-GAPDH (6C5; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-FLAG (M2; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-TBK-1/NAK (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-TBK-1/NAK (Ser172) (D52C2; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-IKKi/IKKε (Abcam, Cambridge, UK), and rabbit anti-phospho-IKKε (Ser172) (D1B7; Cell Signaling Technology, Danvers, MA, USA) were used according to the manufacturers’ instructions.

    Techniques: Activation Assay, Transfection, Expressing, Plasmid Preparation, Activity Assay

    ZIKV NS5 inhibits phosphorylation of IRF3 and IKKε. HEK293 cells were co-transfected with expression plasmids for ( A ) ∆RIG-I (400 ng/well) and wtIRF3 (400 ng/well), ( B ) MAVS (400 ng/well), ( C ) TBK-1 (400 ng/well), ( D ) IKKε (40 ng/well and 30 ng/well) and ( E ) the constitutively active form of IRF3 (IRF3-5D) (400 ng/well), and with expression plasmids for ZIKV NS3 and NS5 (40 ng/well and 400 ng/well, for lower panel on IKKε NS5 plasmid amounts were 200 ng/400 ng/800 ng/1600 ng per well). An expression plasmid for HCV NS3/4A was used as a control. Expression levels and the phosphorylation status of the proteins were detected with immunoblotting.

    Journal: Viruses

    Article Title: Zika Virus Non-Structural Protein NS5 Inhibits the RIG-I Pathway and Interferon Lambda 1 Promoter Activation by Targeting IKK Epsilon

    doi: 10.3390/v11111024

    Figure Lengend Snippet: ZIKV NS5 inhibits phosphorylation of IRF3 and IKKε. HEK293 cells were co-transfected with expression plasmids for ( A ) ∆RIG-I (400 ng/well) and wtIRF3 (400 ng/well), ( B ) MAVS (400 ng/well), ( C ) TBK-1 (400 ng/well), ( D ) IKKε (40 ng/well and 30 ng/well) and ( E ) the constitutively active form of IRF3 (IRF3-5D) (400 ng/well), and with expression plasmids for ZIKV NS3 and NS5 (40 ng/well and 400 ng/well, for lower panel on IKKε NS5 plasmid amounts were 200 ng/400 ng/800 ng/1600 ng per well). An expression plasmid for HCV NS3/4A was used as a control. Expression levels and the phosphorylation status of the proteins were detected with immunoblotting.

    Article Snippet: Antibodies Rabbit anti-IRF3 (FL-425; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-IRF3 (SL-12; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-phospho-IRF3 (Ser396) (4D4G; Cell Signaling Technology, Danvers, MA, USA), mouse anti-HA1.1 Epitope Tag (BioLegend, San Diego, CA, USA), mouse anti-GAPDH (6C5; Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-FLAG (M2; Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-TBK-1/NAK (Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-TBK-1/NAK (Ser172) (D52C2; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-IKKi/IKKε (Abcam, Cambridge, UK), and rabbit anti-phospho-IKKε (Ser172) (D1B7; Cell Signaling Technology, Danvers, MA, USA) were used according to the manufacturers’ instructions.

    Techniques: Transfection, Expressing, Plasmid Preparation

    Neutralization of TLR2 blocks gp120-mediated IFNβ production and IRF3 activation. ( a ) Endometrial GECs were pretreated with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibodies (all at 10 μg/ml) before exposure to gp120 (100 ng/ml) or mock treatment (media). FimH and Pam3CSK4 were used as positive controls for activation of TLR4 and TLR2, respectively. Supernatants were collected after 24 h and analyzed by ELISA for IFNβ production. Data shown are mean + s.d. and representative of three separate experiments done on cells isolated from three different tissues. ( b ) Epithelial monolayers were fixed after 2 h of exposure of gp120 with and without pretreatment with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibody and stained for IRF3. Propidium iodide was used to stain nuclei. Images were captures by a laser-scanning confocal microscopy. Magnification × 1260. Images are representative of one of three separate experiments done on cells isolated from three different tissues. ( c ) Quantitation of IRF3 colocalization were done by Image J software and presented in the graph. Significance was calculated by one-way ANOVA and IRF3 colocalization in all treatments were compared with mock treatment. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Interferon-β induced in female genital epithelium by HIV-1 glycoprotein 120 via Toll-like-receptor 2 pathway acts to protect the mucosal barrier

    doi: 10.1038/cmi.2017.168

    Figure Lengend Snippet: Neutralization of TLR2 blocks gp120-mediated IFNβ production and IRF3 activation. ( a ) Endometrial GECs were pretreated with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibodies (all at 10 μg/ml) before exposure to gp120 (100 ng/ml) or mock treatment (media). FimH and Pam3CSK4 were used as positive controls for activation of TLR4 and TLR2, respectively. Supernatants were collected after 24 h and analyzed by ELISA for IFNβ production. Data shown are mean + s.d. and representative of three separate experiments done on cells isolated from three different tissues. ( b ) Epithelial monolayers were fixed after 2 h of exposure of gp120 with and without pretreatment with neutralizing antibodies against TLR2, TLR4, TLR5 or isotype control antibody and stained for IRF3. Propidium iodide was used to stain nuclei. Images were captures by a laser-scanning confocal microscopy. Magnification × 1260. Images are representative of one of three separate experiments done on cells isolated from three different tissues. ( c ) Quantitation of IRF3 colocalization were done by Image J software and presented in the graph. Significance was calculated by one-way ANOVA and IRF3 colocalization in all treatments were compared with mock treatment. * P

    Article Snippet: Cells were stained with primary mouse monoclonal anti-human IRF3 (SL-12, Santa Cruz Biotechnology, Inc.) and Alexa Flour 488-conjugated secondary antibody, followed by propidium iodide nuclear counterstaining.

    Techniques: Neutralization, Activation Assay, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Confocal Microscopy, Quantitation Assay, Software

    Induction of IFNβ in endometrial GECs by HIV-1 gp120 is mediated through IRF3. Endometrial GECs were exposed to medium or poly I:C, HIV-1 (10 5 IU/well) or gp120 (100 ng/ml alone or with anti-gp120 neutralizing antibody) for 1–3 h. Cells were fixed and stained for the IRF3 (green fluorescence). Nuclear counterstaining (red fluorescence) was achieved using PI. Images were captured by a laser-scanning confocal microscopy. ( a ) Representative images are shown at 2 h time point from one of three separate experiments. Magnification × 1260. ( b ) IRF3 translocation and nuclear colocalization was measured by Image J software and presented as relative light units. ( c ) Time kinetics of IRF3 colocalization following treatment of endometrial GECs with medium or poly I:C (positive control), HIV-1 or gp120. ( d , e ) Endometrial GECs were incubated with the IRF3 inhibitor, BX795 (1 μM) for 1 h, before exposure with gp120, HIV-1 or poly I:C (positive control). Supernatants were collected after 24 h and assayed by ELISA. Results showed IFNβ production in apical ( d ) and basolateral supernatants ( e ). ( f ) Endometrial GECs were treated with BX795 for 1 h before gp120 or HIV-1 exposure for 2 h. The cells were fixed and stained for IRF3 and nuclei. Images were captured by laser-scanning confocal microscopy. Magnification: × 1260. ( g ) Colocalization was measured by image J software and represented in a bar diagram. h Endometrial GECs were preincubated with BX795 or media (mock) for 1 h and TERs were measured pretreatment and after 24 h of treatment with mock or HIV-1 to check whether BX795 was affecting HIV-1-mediated barrier disruption. Images are representatives of three separate experiments from cells isolated from three individual tissues. * P

    Journal: Cellular and Molecular Immunology

    Article Title: Interferon-β induced in female genital epithelium by HIV-1 glycoprotein 120 via Toll-like-receptor 2 pathway acts to protect the mucosal barrier

    doi: 10.1038/cmi.2017.168

    Figure Lengend Snippet: Induction of IFNβ in endometrial GECs by HIV-1 gp120 is mediated through IRF3. Endometrial GECs were exposed to medium or poly I:C, HIV-1 (10 5 IU/well) or gp120 (100 ng/ml alone or with anti-gp120 neutralizing antibody) for 1–3 h. Cells were fixed and stained for the IRF3 (green fluorescence). Nuclear counterstaining (red fluorescence) was achieved using PI. Images were captured by a laser-scanning confocal microscopy. ( a ) Representative images are shown at 2 h time point from one of three separate experiments. Magnification × 1260. ( b ) IRF3 translocation and nuclear colocalization was measured by Image J software and presented as relative light units. ( c ) Time kinetics of IRF3 colocalization following treatment of endometrial GECs with medium or poly I:C (positive control), HIV-1 or gp120. ( d , e ) Endometrial GECs were incubated with the IRF3 inhibitor, BX795 (1 μM) for 1 h, before exposure with gp120, HIV-1 or poly I:C (positive control). Supernatants were collected after 24 h and assayed by ELISA. Results showed IFNβ production in apical ( d ) and basolateral supernatants ( e ). ( f ) Endometrial GECs were treated with BX795 for 1 h before gp120 or HIV-1 exposure for 2 h. The cells were fixed and stained for IRF3 and nuclei. Images were captured by laser-scanning confocal microscopy. Magnification: × 1260. ( g ) Colocalization was measured by image J software and represented in a bar diagram. h Endometrial GECs were preincubated with BX795 or media (mock) for 1 h and TERs were measured pretreatment and after 24 h of treatment with mock or HIV-1 to check whether BX795 was affecting HIV-1-mediated barrier disruption. Images are representatives of three separate experiments from cells isolated from three individual tissues. * P

    Article Snippet: Cells were stained with primary mouse monoclonal anti-human IRF3 (SL-12, Santa Cruz Biotechnology, Inc.) and Alexa Flour 488-conjugated secondary antibody, followed by propidium iodide nuclear counterstaining.

    Techniques: Staining, Fluorescence, Confocal Microscopy, Translocation Assay, Software, Positive Control, Incubation, Enzyme-linked Immunosorbent Assay, Isolation

    R6 competes with IRF3 for binding to CBP. (A) Telomerized RFs were transfected with pcDNA3.1-R6HA (5, 10, or 20 μg DNA) or empty pcDNA3.1 for 40 h and subsequently transfected with poly(I · C) for 6 h. Nuclear lysates were immunoprecipitated

    Journal: Journal of Virology

    Article Title: A Rhesus Rhadinovirus Viral Interferon (IFN) Regulatory Factor Is Virion Associated and Inhibits the Early IFN Antiviral Response

    doi: 10.1128/JVI.01175-15

    Figure Lengend Snippet: R6 competes with IRF3 for binding to CBP. (A) Telomerized RFs were transfected with pcDNA3.1-R6HA (5, 10, or 20 μg DNA) or empty pcDNA3.1 for 40 h and subsequently transfected with poly(I · C) for 6 h. Nuclear lysates were immunoprecipitated

    Article Snippet: To demonstrate the specificity of protein-DNA complex formation, 500-fold molar excess of unlabeled wild-type oligonucleotide corresponding to the PRDI-PRDIII region of the IFN-β promoter was added before adding labeled probe or was preincubated with anti-CBP (H100; Santa Cruz) or anti-IRF3 (SL-12; Santa Cruz).

    Techniques: Binding Assay, Transfection, Immunoprecipitation

    Potential model of IFN inhibition by R6. Upon detection of RRV infection by TLR3, RIG-I, or MDA-5, TBK1 is activated and subsequently phosphorylates IRF3. pIRF3 then dimerizes and translocates to the nucleus. Within the nucleus, R6 binds to the transcriptional

    Journal: Journal of Virology

    Article Title: A Rhesus Rhadinovirus Viral Interferon (IFN) Regulatory Factor Is Virion Associated and Inhibits the Early IFN Antiviral Response

    doi: 10.1128/JVI.01175-15

    Figure Lengend Snippet: Potential model of IFN inhibition by R6. Upon detection of RRV infection by TLR3, RIG-I, or MDA-5, TBK1 is activated and subsequently phosphorylates IRF3. pIRF3 then dimerizes and translocates to the nucleus. Within the nucleus, R6 binds to the transcriptional

    Article Snippet: To demonstrate the specificity of protein-DNA complex formation, 500-fold molar excess of unlabeled wild-type oligonucleotide corresponding to the PRDI-PRDIII region of the IFN-β promoter was added before adding labeled probe or was preincubated with anti-CBP (H100; Santa Cruz) or anti-IRF3 (SL-12; Santa Cruz).

    Techniques: Inhibition, Infection, Multiple Displacement Amplification