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  • 86
    Millipore stat1 β β mice
    STAT1β shows prolonged tyrosine 701 phosphorylation. (A to D) BMMϕ derived from WT (+/+), <t>Stat1</t> <t>β/β</t> (β/β), and Stat1 α/α (α/α) mice were stimulated with IFN-β (A) or
    Stat1 β β Mice, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc stat1
    Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of <t>Stat1,</t> phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P
    Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 4691 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stat1/product/Cell Signaling Technology Inc
    Average 99 stars, based on 4691 article reviews
    Price from $9.99 to $1999.99
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    91
    R&D Systems stat1 inhibitor
    Pharmacological inhibition of both PTP1B and TC-PTP with specific inhibitor. (A) Specificity of 1B/TC inh (50 μM) to inhibit PTP1B and TC-PTP dephosphorylation ( n = 3). (B) Titration of 1B/TC inh in mouse moDCs using IL-12 production as indicator of activation ( n = 3). (C) IFNγ production by 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 4). (D) Expression levels (MFI) of CD40, CD80, CD86, MHC class I and II on 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 3). (E) Activation status <t>STAT1</t> and STAT4 ( n = 3). (F) Activation status of Src kinase and IkBα ( n = 4). Therapeutic properties of 1B/TC inh-treated moDCs in a mouse model of E.G7 lymphoma: (G) Tumor volume before moDC treatment (10 d after implantation of 5 × 10 5 E.G7-OVA cells) and (H) after intraperitoneal (IP) injections of 5 × 10 6 1B/TC inh-treated or non-treated moDCs compared with control group (tumor-bearing mice without moDC treatments). The IP injections with moDCs were given 18 d after tumor implantation. (I) Images represent tumor-bearing mice 8 d after of moDC treatment. The results are representative of at least three independent experiments. The differences in more than two groups were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric. The differences between two groups were determined with unpaired t test (two tails of distribution). Significant differences are represented by p values
    Stat1 Inhibitor, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc phospho stat1
    Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of <t>Stat1,</t> phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P
    Phospho Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 933 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho stat1/product/Cell Signaling Technology Inc
    Average 99 stars, based on 933 article reviews
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    96
    Selleck Chemicals stat1 inhibitor fludarabine
    IFNγ induces MPC1/2 downregulation via stat3 activation. (A) After treated MC38 and CaCO-2 cells with PBS or IFNγ (50 ng/ml) for 48 h, the cell lysates were extracted and the levels of phosphorylation <t>STAT1,</t> phosphorylation STAT3 STAT1 and STAT3 were detected with western-blot, β-actin was used as internal control. (B-C) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with <t>Fludarabine</t> (1 uM) or IFNγ combined with Stattic (5 uM) for 48 h. (B) The cell lysates were extracted and the levels of MPC1 and MPC2 were detected with western-blot. (C) 1.5 × 10 4 of treated cells were seeded in 96-well plate in 100ul medium. 24 h later the supernants were collected and the level of lactate was detected. (D) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (5uM) for 48 h. the cells were trypsinized and stained with CellROX™ and detected the overall ROS level with flow cytometry. (E) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (10 uM) for 48 h. The cell lysates were extracted and the levels of full lenghth Caspase 7 and Caspase 3 and cleaved Caspase 7 and cleaved Caspase 3 were detected by western-blot and β-actin was used as internal control. In the same time, the cells were trypsinized and stained with Annexin-V and 7-AAD. The overall apoptosis was detected with flow cytometry. (F) Mice were inoculated with 5 × 10 5 MC38 cells. When tumor size was 5 × 5 mm, mice were treated with PBS, IFNγ (10 μg/mouse), Stattic (10 mg/kg) or IFN-γ combined with Stattic intratumorally for 10 days. Tumor growth was measured (n = 6). Data shown are representative of three independent experiments and error bars represent mean ± s.e.m., N.S., no significant difference; *p
    Stat1 Inhibitor Fludarabine, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 96/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cell Signaling Technology Inc phospho stat1α β
    Autophagy was important for IFN-γ-activated STAT1. A , after IFN-γ (10 ng/ml) treatment, Western blotting was used to determine the time kinetic phosphorylation of <t>STAT1α/β</t> (Tyr 701 ) as well as IRF1 and SOCS1 expression in
    Phospho Stat1α β, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    STAT1β shows prolonged tyrosine 701 phosphorylation. (A to D) BMMϕ derived from WT (+/+), Stat1 β/β (β/β), and Stat1 α/α (α/α) mice were stimulated with IFN-β (A) or

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: STAT1β shows prolonged tyrosine 701 phosphorylation. (A to D) BMMϕ derived from WT (+/+), Stat1 β/β (β/β), and Stat1 α/α (α/α) mice were stimulated with IFN-β (A) or

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: Derivative Assay, Mouse Assay

    STAT1β shows prolonged nuclear localization and prolonged promoter binding after IFN-γ treatment compared to STAT1α. (A) BMMϕ derived from WT (+/+), Stat1 β/β (β/β), and Stat1 α/α

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: STAT1β shows prolonged nuclear localization and prolonged promoter binding after IFN-γ treatment compared to STAT1α. (A) BMMϕ derived from WT (+/+), Stat1 β/β (β/β), and Stat1 α/α

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: Binding Assay, Derivative Assay

    STAT1α and STAT1β can mediate type I and type III IFN-dependent antiviral immunity in vivo . (A) EMCV (50 PFU/mouse) was administered i.p. to WT ( Stat1 +/+ ), Stat1 β/β , Stat1 α/α , and Stat1 −/−

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: STAT1α and STAT1β can mediate type I and type III IFN-dependent antiviral immunity in vivo . (A) EMCV (50 PFU/mouse) was administered i.p. to WT ( Stat1 +/+ ), Stat1 β/β , Stat1 α/α , and Stat1 −/−

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: In Vivo

    STAT1β is transcriptionally active in response to IFN-β and IFN-γ. BMMϕ isolated from WT (+/+), Stat1 β/β (β/β), Stat1 α/α (α/α), and Stat1 −/−

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: STAT1β is transcriptionally active in response to IFN-β and IFN-γ. BMMϕ isolated from WT (+/+), Stat1 β/β (β/β), Stat1 α/α (α/α), and Stat1 −/−

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: Isolation

    STAT1α and STAT1β show differential efficiencies in immune defense against MCMV and L. monocytogenes infections. (A) WT ( Stat1 +/+ ), Stat1 β/β , Stat1 α/α , and Stat1 −/− mice were infected i.p.

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: STAT1α and STAT1β show differential efficiencies in immune defense against MCMV and L. monocytogenes infections. (A) WT ( Stat1 +/+ ), Stat1 β/β , Stat1 α/α , and Stat1 −/− mice were infected i.p.

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: Mouse Assay, Infection

    Transcriptional activities of STAT1α and STAT1β overlap but are nonredundant. BMMϕ isolated from WT (+/+), Stat1 β/β (β/β), Stat1 α/α (α/α), and Stat1 −/−

    Journal: Molecular and Cellular Biology

    Article Title: STAT1β Is Not Dominant Negative and Is Capable of Contributing to Gamma Interferon-Dependent Innate Immunity

    doi: 10.1128/MCB.00295-14

    Figure Lengend Snippet: Transcriptional activities of STAT1α and STAT1β overlap but are nonredundant. BMMϕ isolated from WT (+/+), Stat1 β/β (β/β), Stat1 α/α (α/α), and Stat1 −/−

    Article Snippet: Genetic screening of WT, Stat1 α/α , and Stat1 β/β mice was performed by a duplex PCR with primers (purchased from Sigma-Aldrich) using DNA from mouse tail biopsy specimens.

    Techniques: Isolation

    Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of Stat1, phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P

    Journal: PLoS ONE

    Article Title: Cardiac-Specific Activation of IKK2 Leads to Defects in Heart Development and Embryonic Lethality

    doi: 10.1371/journal.pone.0141591

    Figure Lengend Snippet: Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of Stat1, phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P

    Article Snippet: Primary antibodies against the following antigens were used: human IKK2 (Abcam Y466), IKK1/2 (Santa Cruz sc-7607, lot H2208), ERK2 (Santa Cruz sc-154, lot F0210), p21 (Santa Cruz sc-6246, lot A1209), Stat1 (Cell Signaling 9172, lot 14), phospho-Stat1 (Cell Signaling 9167, lot 6), and luciferase (Promega G7451).

    Techniques: Expressing, Luciferase, Western Blot, Real-time Polymerase Chain Reaction, Purification

    Pharmacological inhibition of both PTP1B and TC-PTP with specific inhibitor. (A) Specificity of 1B/TC inh (50 μM) to inhibit PTP1B and TC-PTP dephosphorylation ( n = 3). (B) Titration of 1B/TC inh in mouse moDCs using IL-12 production as indicator of activation ( n = 3). (C) IFNγ production by 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 4). (D) Expression levels (MFI) of CD40, CD80, CD86, MHC class I and II on 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 3). (E) Activation status STAT1 and STAT4 ( n = 3). (F) Activation status of Src kinase and IkBα ( n = 4). Therapeutic properties of 1B/TC inh-treated moDCs in a mouse model of E.G7 lymphoma: (G) Tumor volume before moDC treatment (10 d after implantation of 5 × 10 5 E.G7-OVA cells) and (H) after intraperitoneal (IP) injections of 5 × 10 6 1B/TC inh-treated or non-treated moDCs compared with control group (tumor-bearing mice without moDC treatments). The IP injections with moDCs were given 18 d after tumor implantation. (I) Images represent tumor-bearing mice 8 d after of moDC treatment. The results are representative of at least three independent experiments. The differences in more than two groups were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric. The differences between two groups were determined with unpaired t test (two tails of distribution). Significant differences are represented by p values

    Journal: Oncoimmunology

    Article Title: Downregulation of PTP1B and TC-PTP phosphatases potentiate dendritic cell-based immunotherapy through IL-12/IFNγ signaling

    doi: 10.1080/2162402X.2017.1321185

    Figure Lengend Snippet: Pharmacological inhibition of both PTP1B and TC-PTP with specific inhibitor. (A) Specificity of 1B/TC inh (50 μM) to inhibit PTP1B and TC-PTP dephosphorylation ( n = 3). (B) Titration of 1B/TC inh in mouse moDCs using IL-12 production as indicator of activation ( n = 3). (C) IFNγ production by 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 4). (D) Expression levels (MFI) of CD40, CD80, CD86, MHC class I and II on 1B/TC inh-treated mature moDCs (4 μM dose) ( n = 3). (E) Activation status STAT1 and STAT4 ( n = 3). (F) Activation status of Src kinase and IkBα ( n = 4). Therapeutic properties of 1B/TC inh-treated moDCs in a mouse model of E.G7 lymphoma: (G) Tumor volume before moDC treatment (10 d after implantation of 5 × 10 5 E.G7-OVA cells) and (H) after intraperitoneal (IP) injections of 5 × 10 6 1B/TC inh-treated or non-treated moDCs compared with control group (tumor-bearing mice without moDC treatments). The IP injections with moDCs were given 18 d after tumor implantation. (I) Images represent tumor-bearing mice 8 d after of moDC treatment. The results are representative of at least three independent experiments. The differences in more than two groups were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric. The differences between two groups were determined with unpaired t test (two tails of distribution). Significant differences are represented by p values

    Article Snippet: STAT1 and STAT4 inhibition Mouse PTP-deficient moDCs were generated as described previously and maturation was induced for additional 24 h in the presence of LPS and 50 μM of STAT1 inhibitor (Fludarabine, R & D Systems, cat# 3495) or 50 μM of STAT4 inhibitor (Lisofyllin, Santa Cruz Biotechnology, cat# sc-201055).

    Techniques: Inhibition, De-Phosphorylation Assay, Titration, Activation Assay, Expressing, Mouse Assay, Tumor Implantation

    Application of 1B/TC inh-treated human DCs in cancer immunotherapy. (A) Activation status of STAT1 and STAT4 in 1B/TC inh-treated human moDCs derived from a pancreatic cancer (PaC) patient ( n = 3). (B) IL-12 production by 1B/TC inh-treated human moDCs from four PaC patients. (C) Frequency of antigen-specific (CEA and CA19–9) activated T cells measured based on IFNγ production detected in ELISpot assay. The differences in more than two groups were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric. The differences between two groups were determined with unpaired t -test (two tails of distribution). Significant differences are represented by p values

    Journal: Oncoimmunology

    Article Title: Downregulation of PTP1B and TC-PTP phosphatases potentiate dendritic cell-based immunotherapy through IL-12/IFNγ signaling

    doi: 10.1080/2162402X.2017.1321185

    Figure Lengend Snippet: Application of 1B/TC inh-treated human DCs in cancer immunotherapy. (A) Activation status of STAT1 and STAT4 in 1B/TC inh-treated human moDCs derived from a pancreatic cancer (PaC) patient ( n = 3). (B) IL-12 production by 1B/TC inh-treated human moDCs from four PaC patients. (C) Frequency of antigen-specific (CEA and CA19–9) activated T cells measured based on IFNγ production detected in ELISpot assay. The differences in more than two groups were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric. The differences between two groups were determined with unpaired t -test (two tails of distribution). Significant differences are represented by p values

    Article Snippet: STAT1 and STAT4 inhibition Mouse PTP-deficient moDCs were generated as described previously and maturation was induced for additional 24 h in the presence of LPS and 50 μM of STAT1 inhibitor (Fludarabine, R & D Systems, cat# 3495) or 50 μM of STAT4 inhibitor (Lisofyllin, Santa Cruz Biotechnology, cat# sc-201055).

    Techniques: Activation Assay, Derivative Assay, Enzyme-linked Immunospot

    Simultaneous downregulation of PTP1B and TC-PTP enhances moDCs maturation. (A) Comparisons between double heterozygous (1B:TC fl/wt ) and TC fl/wt moDCs, 1B fl/wt moDCs and wt moDCs respectively. MFI values of the expression of MHC class I, CD86 and CD80 ( n = 5). (B) Production of Th1 polarizing cytokines ( n = 5): (B) IL-12 and (C) IFNγ. (D) Quantification of IFNγ produced by activated OT-I CD8 + T cells co-cultured with mature and OVA-pulsed 1B:TC fl/wt, 1B fl/wt , TC fl/wt or wt moDCs for 48 h ( n = 8–9). (E) Quantification of IFNγ produced by activated OT-II CD4 + T cells co-cultured with mature and OVA-pulsed 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs for 48 h ( n = 5). (F) Activation status of Src kinase and IkBa downstream of TLR4 ( n = 3). Significant differences are represented by p values and (*) indicate significant differences between wt DCs and the rest of the groups. (G) Activation status of STAT1, STAT4 and Src in STAT1 inhibitor (fludarabine) or STAT4 inhibitor (lisofylline) treated or non-treated 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs ( n = 3). Production of Th1-polarizing cytokines by 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs previously treated with STAT1- or STAT4-inhibitor during the maturation process ( n = 3): (H) IL-12 and (I) IFNγ. The results are representative of at least three independent experiments. Significant differences among the groups are represented by p values and (*) indicate significant differences within a group. The comparisons were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric.

    Journal: Oncoimmunology

    Article Title: Downregulation of PTP1B and TC-PTP phosphatases potentiate dendritic cell-based immunotherapy through IL-12/IFNγ signaling

    doi: 10.1080/2162402X.2017.1321185

    Figure Lengend Snippet: Simultaneous downregulation of PTP1B and TC-PTP enhances moDCs maturation. (A) Comparisons between double heterozygous (1B:TC fl/wt ) and TC fl/wt moDCs, 1B fl/wt moDCs and wt moDCs respectively. MFI values of the expression of MHC class I, CD86 and CD80 ( n = 5). (B) Production of Th1 polarizing cytokines ( n = 5): (B) IL-12 and (C) IFNγ. (D) Quantification of IFNγ produced by activated OT-I CD8 + T cells co-cultured with mature and OVA-pulsed 1B:TC fl/wt, 1B fl/wt , TC fl/wt or wt moDCs for 48 h ( n = 8–9). (E) Quantification of IFNγ produced by activated OT-II CD4 + T cells co-cultured with mature and OVA-pulsed 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs for 48 h ( n = 5). (F) Activation status of Src kinase and IkBa downstream of TLR4 ( n = 3). Significant differences are represented by p values and (*) indicate significant differences between wt DCs and the rest of the groups. (G) Activation status of STAT1, STAT4 and Src in STAT1 inhibitor (fludarabine) or STAT4 inhibitor (lisofylline) treated or non-treated 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs ( n = 3). Production of Th1-polarizing cytokines by 1B:TC fl/wt , 1B fl/wt , TC fl/wt or wt moDCs previously treated with STAT1- or STAT4-inhibitor during the maturation process ( n = 3): (H) IL-12 and (I) IFNγ. The results are representative of at least three independent experiments. Significant differences among the groups are represented by p values and (*) indicate significant differences within a group. The comparisons were determined using One-Way ANOVA (Holm–Sidak multiple comparison test) for parametric and Dunn's multiple test for non-parametric.

    Article Snippet: STAT1 and STAT4 inhibition Mouse PTP-deficient moDCs were generated as described previously and maturation was induced for additional 24 h in the presence of LPS and 50 μM of STAT1 inhibitor (Fludarabine, R & D Systems, cat# 3495) or 50 μM of STAT4 inhibitor (Lisofyllin, Santa Cruz Biotechnology, cat# sc-201055).

    Techniques: Expressing, Produced, Cell Culture, Activation Assay

    Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of Stat1, phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P

    Journal: PLoS ONE

    Article Title: Cardiac-Specific Activation of IKK2 Leads to Defects in Heart Development and Embryonic Lethality

    doi: 10.1371/journal.pone.0141591

    Figure Lengend Snippet: Dysregulation of regulators of proliferation and differentiation in embryonic IKK MyHC hearts. (A) The expression of Stat1, phospho-Stat1, p21 and the transgenes IKK2 and luciferase was determined by Western blot, with Erk2 shown as loading control. The diagram on the right shows a quantification of the p21 immunoblot, normalized to the expression of ERK2. (B) Expression of the indicated cell cycle modulators was determined at the mRNA level using real-time quantitative PCR in purified cardiomyocytes from IKK MyHC (black bars) and control (grey bars) embryos at E10.5. Shown are the means +SEM; N = 5, *P

    Article Snippet: Primary antibodies against the following antigens were used: human IKK2 (Abcam Y466), IKK1/2 (Santa Cruz sc-7607, lot H2208), ERK2 (Santa Cruz sc-154, lot F0210), p21 (Santa Cruz sc-6246, lot A1209), Stat1 (Cell Signaling 9172, lot 14), phospho-Stat1 (Cell Signaling 9167, lot 6), and luciferase (Promega G7451).

    Techniques: Expressing, Luciferase, Western Blot, Real-time Polymerase Chain Reaction, Purification

    IFNγ induces MPC1/2 downregulation via stat3 activation. (A) After treated MC38 and CaCO-2 cells with PBS or IFNγ (50 ng/ml) for 48 h, the cell lysates were extracted and the levels of phosphorylation STAT1, phosphorylation STAT3 STAT1 and STAT3 were detected with western-blot, β-actin was used as internal control. (B-C) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Fludarabine (1 uM) or IFNγ combined with Stattic (5 uM) for 48 h. (B) The cell lysates were extracted and the levels of MPC1 and MPC2 were detected with western-blot. (C) 1.5 × 10 4 of treated cells were seeded in 96-well plate in 100ul medium. 24 h later the supernants were collected and the level of lactate was detected. (D) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (5uM) for 48 h. the cells were trypsinized and stained with CellROX™ and detected the overall ROS level with flow cytometry. (E) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (10 uM) for 48 h. The cell lysates were extracted and the levels of full lenghth Caspase 7 and Caspase 3 and cleaved Caspase 7 and cleaved Caspase 3 were detected by western-blot and β-actin was used as internal control. In the same time, the cells were trypsinized and stained with Annexin-V and 7-AAD. The overall apoptosis was detected with flow cytometry. (F) Mice were inoculated with 5 × 10 5 MC38 cells. When tumor size was 5 × 5 mm, mice were treated with PBS, IFNγ (10 μg/mouse), Stattic (10 mg/kg) or IFN-γ combined with Stattic intratumorally for 10 days. Tumor growth was measured (n = 6). Data shown are representative of three independent experiments and error bars represent mean ± s.e.m., N.S., no significant difference; *p

    Journal: Redox Biology

    Article Title: Enhanced mitochondrial pyruvate transport elicits a robust ROS production to sensitize the antitumor efficacy of interferon-γ in colon cancer

    doi: 10.1016/j.redox.2018.10.024

    Figure Lengend Snippet: IFNγ induces MPC1/2 downregulation via stat3 activation. (A) After treated MC38 and CaCO-2 cells with PBS or IFNγ (50 ng/ml) for 48 h, the cell lysates were extracted and the levels of phosphorylation STAT1, phosphorylation STAT3 STAT1 and STAT3 were detected with western-blot, β-actin was used as internal control. (B-C) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Fludarabine (1 uM) or IFNγ combined with Stattic (5 uM) for 48 h. (B) The cell lysates were extracted and the levels of MPC1 and MPC2 were detected with western-blot. (C) 1.5 × 10 4 of treated cells were seeded in 96-well plate in 100ul medium. 24 h later the supernants were collected and the level of lactate was detected. (D) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (5uM) for 48 h. the cells were trypsinized and stained with CellROX™ and detected the overall ROS level with flow cytometry. (E) After treated MC38 and CaCO-2 cells with IFNγ (50 ng/ml) or IFNγ combined with Stattic (10 uM) for 48 h. The cell lysates were extracted and the levels of full lenghth Caspase 7 and Caspase 3 and cleaved Caspase 7 and cleaved Caspase 3 were detected by western-blot and β-actin was used as internal control. In the same time, the cells were trypsinized and stained with Annexin-V and 7-AAD. The overall apoptosis was detected with flow cytometry. (F) Mice were inoculated with 5 × 10 5 MC38 cells. When tumor size was 5 × 5 mm, mice were treated with PBS, IFNγ (10 μg/mouse), Stattic (10 mg/kg) or IFN-γ combined with Stattic intratumorally for 10 days. Tumor growth was measured (n = 6). Data shown are representative of three independent experiments and error bars represent mean ± s.e.m., N.S., no significant difference; *p

    Article Snippet: STAT1 inhibitor Fludarabine and STAT3 inhibitor Stattic were purchased from Selleck Chemicals.

    Techniques: Activation Assay, Western Blot, Staining, Flow Cytometry, Cytometry, Mouse Assay

    Autophagy was important for IFN-γ-activated STAT1. A , after IFN-γ (10 ng/ml) treatment, Western blotting was used to determine the time kinetic phosphorylation of STAT1α/β (Tyr 701 ) as well as IRF1 and SOCS1 expression in

    Journal: The Journal of Biological Chemistry

    Article Title: Autophagy Facilitates IFN-?-induced Jak2-STAT1 Activation and Cellular Inflammation *

    doi: 10.1074/jbc.M110.133355

    Figure Lengend Snippet: Autophagy was important for IFN-γ-activated STAT1. A , after IFN-γ (10 ng/ml) treatment, Western blotting was used to determine the time kinetic phosphorylation of STAT1α/β (Tyr 701 ) as well as IRF1 and SOCS1 expression in

    Article Snippet: Antibodies against phospho-Pyk2 (Tyr402 ), Pyk2, phospho-STAT1α/β (Tyr701 ), STAT1α/β, phospho-Jak2 (Tyr1007 /Tyr1008 ), Jak2, phospho-Jak1 (Tyr1022 /Tyr1023 ), Jak1, IFN regulatory factor-1 (IRF1), SOCS1, SHP2, and COX IV were purchased from Cell Signaling Technology.

    Techniques: Western Blot, Expressing

    Autophagy was required for IFN-γ-activated Jak2-STAT1 and inflammation. Western blotting was used to determine LC3 conversion ( A ) and the time kinetic phosphorylation of Jak2 (Tyr 1007 /Tyr 1008 ) and STAT1α/β (Tyr 701 ) ( B ) in IFN-γ

    Journal: The Journal of Biological Chemistry

    Article Title: Autophagy Facilitates IFN-?-induced Jak2-STAT1 Activation and Cellular Inflammation *

    doi: 10.1074/jbc.M110.133355

    Figure Lengend Snippet: Autophagy was required for IFN-γ-activated Jak2-STAT1 and inflammation. Western blotting was used to determine LC3 conversion ( A ) and the time kinetic phosphorylation of Jak2 (Tyr 1007 /Tyr 1008 ) and STAT1α/β (Tyr 701 ) ( B ) in IFN-γ

    Article Snippet: Antibodies against phospho-Pyk2 (Tyr402 ), Pyk2, phospho-STAT1α/β (Tyr701 ), STAT1α/β, phospho-Jak2 (Tyr1007 /Tyr1008 ), Jak2, phospho-Jak1 (Tyr1022 /Tyr1023 ), Jak1, IFN regulatory factor-1 (IRF1), SOCS1, SHP2, and COX IV were purchased from Cell Signaling Technology.

    Techniques: Western Blot

    ROS-regulated SHP2 inhibited IFN-γ-activated STAT1 in the absence of autophagy. A , Western blotting was used to determine IFN-γ (10 ng/ml)-induced phosphorylation of STAT1α/β (Tyr 701 ) in WT MEFs 0.25 h after they had been

    Journal: The Journal of Biological Chemistry

    Article Title: Autophagy Facilitates IFN-?-induced Jak2-STAT1 Activation and Cellular Inflammation *

    doi: 10.1074/jbc.M110.133355

    Figure Lengend Snippet: ROS-regulated SHP2 inhibited IFN-γ-activated STAT1 in the absence of autophagy. A , Western blotting was used to determine IFN-γ (10 ng/ml)-induced phosphorylation of STAT1α/β (Tyr 701 ) in WT MEFs 0.25 h after they had been

    Article Snippet: Antibodies against phospho-Pyk2 (Tyr402 ), Pyk2, phospho-STAT1α/β (Tyr701 ), STAT1α/β, phospho-Jak2 (Tyr1007 /Tyr1008 ), Jak2, phospho-Jak1 (Tyr1022 /Tyr1023 ), Jak1, IFN regulatory factor-1 (IRF1), SOCS1, SHP2, and COX IV were purchased from Cell Signaling Technology.

    Techniques: Western Blot