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    Name:
    Stat1 Control Cell Extracts
    Description:
    Nonphosphorylated Stat1 Control Cell Extracts Total cell extracts from HeLa cells prepared without treatment serve as a negative control Supplied in SDS Sample Buffer
    Catalog Number:
    9173
    Price:
    None
    Category:
    Experimental Controls
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    Structured Review

    Cell Signaling Technology Inc stat1
    Inflammatory responses of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were collected 24 h after the injection for real-time RT-PCR and immunoblot analyses. Real-time RT-PCR analyses-based determination of CCL2 ( A ), CCL3 ( B ), CXCL1 ( C ), ICAM-1 ( D ), IL-1β ( E ), IL-10 ( F ), and TNF-α ( G ) mRNA-expression levels. Determination of phosphorylated signal transducer and activator of transcription (p-STAT)1, <t>STAT1,</t> p-STAT3, and STAT3 protein levels by immunoblotting ( H ). Quantification and normalization of p-STAT1 ( I ) and p-STAT3 ( J ) band intensities to those of STAT1 and STAT3, respectively. The data shown are representative of two independent experiments. Values are presented as means ± SD ( n = 6/group). For mRNA expression, significant differences between LPS-treated animals in the saccular phase versus alveolar phase of lung development are indicated by *** P  
    Nonphosphorylated Stat1 Control Cell Extracts Total cell extracts from HeLa cells prepared without treatment serve as a negative control Supplied in SDS Sample Buffer
    https://www.bioz.com/result/stat1/product/Cell Signaling Technology Inc
    Average 99 stars, based on 512 article reviews
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    stat1 - by Bioz Stars, 2020-11
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    Images

    1) Product Images from "Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice"

    Article Title: Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00560.2017

    Inflammatory responses of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were collected 24 h after the injection for real-time RT-PCR and immunoblot analyses. Real-time RT-PCR analyses-based determination of CCL2 ( A ), CCL3 ( B ), CXCL1 ( C ), ICAM-1 ( D ), IL-1β ( E ), IL-10 ( F ), and TNF-α ( G ) mRNA-expression levels. Determination of phosphorylated signal transducer and activator of transcription (p-STAT)1, STAT1, p-STAT3, and STAT3 protein levels by immunoblotting ( H ). Quantification and normalization of p-STAT1 ( I ) and p-STAT3 ( J ) band intensities to those of STAT1 and STAT3, respectively. The data shown are representative of two independent experiments. Values are presented as means ± SD ( n = 6/group). For mRNA expression, significant differences between LPS-treated animals in the saccular phase versus alveolar phase of lung development are indicated by *** P  
    Figure Legend Snippet: Inflammatory responses of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were collected 24 h after the injection for real-time RT-PCR and immunoblot analyses. Real-time RT-PCR analyses-based determination of CCL2 ( A ), CCL3 ( B ), CXCL1 ( C ), ICAM-1 ( D ), IL-1β ( E ), IL-10 ( F ), and TNF-α ( G ) mRNA-expression levels. Determination of phosphorylated signal transducer and activator of transcription (p-STAT)1, STAT1, p-STAT3, and STAT3 protein levels by immunoblotting ( H ). Quantification and normalization of p-STAT1 ( I ) and p-STAT3 ( J ) band intensities to those of STAT1 and STAT3, respectively. The data shown are representative of two independent experiments. Values are presented as means ± SD ( n = 6/group). For mRNA expression, significant differences between LPS-treated animals in the saccular phase versus alveolar phase of lung development are indicated by *** P  

    Techniques Used: Mouse Assay, Injection, Quantitative RT-PCR, Expressing

    Validation of the reverse-phase protein array results in immunoblot assays. The lung tissues of 4-day-old mice were collected 24 h after treatment with the vehicle control (PBS) or one 10 mg/kg dose of LPS (L10) and analyzed in immunoblot assays to determine the protein expression levels of fibroblast growth factor receptor 1 (FGFR1), signal transducer and activator of transcription (STAT)1, STAT3, and annexin-1 ( A ). Densitometric analyses wherein FGFR1 ( B ), STAT1 ( C ), STAT3 ( D ), and annexin-1 ( E ) band intensities were quantified and normalized to β-actin. Values shown are presented as means ± SD ( n = 4/group). Significant differences between PBS- and L10-exposed animals are indicated by *** P
    Figure Legend Snippet: Validation of the reverse-phase protein array results in immunoblot assays. The lung tissues of 4-day-old mice were collected 24 h after treatment with the vehicle control (PBS) or one 10 mg/kg dose of LPS (L10) and analyzed in immunoblot assays to determine the protein expression levels of fibroblast growth factor receptor 1 (FGFR1), signal transducer and activator of transcription (STAT)1, STAT3, and annexin-1 ( A ). Densitometric analyses wherein FGFR1 ( B ), STAT1 ( C ), STAT3 ( D ), and annexin-1 ( E ) band intensities were quantified and normalized to β-actin. Values shown are presented as means ± SD ( n = 4/group). Significant differences between PBS- and L10-exposed animals are indicated by *** P

    Techniques Used: Protein Array, Mouse Assay, Expressing

    2) Product Images from "Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3"

    Article Title: Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1193-6

    VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P
    Figure Legend Snippet: VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P

    Techniques Used: Expressing

    VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P
    Figure Legend Snippet: VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P

    Techniques Used: Expressing, Co-Immunoprecipitation Assay

    3) Product Images from "Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3"

    Article Title: Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1193-6

    VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P
    Figure Legend Snippet: VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P

    Techniques Used: Expressing

    VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P
    Figure Legend Snippet: VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P

    Techniques Used: Expressing, Co-Immunoprecipitation Assay

    4) Product Images from "Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3"

    Article Title: Valproic acid attenuates traumatic spinal cord injury-induced inflammation via STAT1 and NF-κB pathway dependent of HDAC3

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1193-6

    VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P
    Figure Legend Snippet: VPA treatment elevates STAT1 expression in the lesioned spinal cord 7 days after SCI. a STAT1 expressions were higher in the SCI than in the sham group. The STAT1 protein levels were upregulated following the VPA treatment ( P

    Techniques Used: Expressing

    VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P
    Figure Legend Snippet: VPA treatment suppressed the NF-κB pathway via elevating STAT1 expression in the lesioned spinal cord 7 days after SCI. a Co-IP analysis showed a greater elevation in the STAT1 after the VPA treatment than in the SCI group ( P

    Techniques Used: Expressing, Co-Immunoprecipitation Assay

    5) Product Images from "miR-29c plays a suppressive role in breast cancer by targeting the TIMP3/STAT1/FOXO1 pathway"

    Article Title: miR-29c plays a suppressive role in breast cancer by targeting the TIMP3/STAT1/FOXO1 pathway

    Journal: Clinical Epigenetics

    doi: 10.1186/s13148-018-0495-y

    miR-29c directly targeted DNMT3B and regulated the DNMT3B/TIMP3/STAT1/FOXO1 pathway in breast cancer cells. a The potential binding site of miR-29c in the 3′UTR of DNMT3B. b Dual-Luciferase Reporter Assay of miR-29c and DNMT3B in MDA-MB-231 cells. c Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MCF-7 cells after the transfection of miR-29c inhibitor. d qMS-PCR assay of the methylation level of TIMP3 in MCF-7 cells after the transfection of miR-29c inhibitor. e Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MDA-MB-231 cells after the transfection of miR-29c mimic. f qMS-PCR assay of the methylation level of TIMP3 in MDA-MB-231 cells after the transfection of miR-29c mimic. g Migration assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. h Invasion assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. i Protein levels of STAT1 and FOXO1 detected by Western blotting in MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. Data are presented as mean ± SD from three independent experiments, and every experiment was repeated three times, * P
    Figure Legend Snippet: miR-29c directly targeted DNMT3B and regulated the DNMT3B/TIMP3/STAT1/FOXO1 pathway in breast cancer cells. a The potential binding site of miR-29c in the 3′UTR of DNMT3B. b Dual-Luciferase Reporter Assay of miR-29c and DNMT3B in MDA-MB-231 cells. c Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MCF-7 cells after the transfection of miR-29c inhibitor. d qMS-PCR assay of the methylation level of TIMP3 in MCF-7 cells after the transfection of miR-29c inhibitor. e Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MDA-MB-231 cells after the transfection of miR-29c mimic. f qMS-PCR assay of the methylation level of TIMP3 in MDA-MB-231 cells after the transfection of miR-29c mimic. g Migration assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. h Invasion assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. i Protein levels of STAT1 and FOXO1 detected by Western blotting in MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. Data are presented as mean ± SD from three independent experiments, and every experiment was repeated three times, * P

    Techniques Used: Binding Assay, Luciferase, Reporter Assay, Multiple Displacement Amplification, Western Blot, Transfection, Polymerase Chain Reaction, Methylation, Migration

    Proposed mechanistic scheme: miR-29c suppresses breast cancer by the TIMP3/STAT1/FOXO1 pathway. The expression of miR-29c was decreased in breast cancer, which leading to the elevated expression of DNMT3B that is critical for promoter methylation and decreases the expression of TIMP3. As a result, the expression of STAT1 was increased, and it inhibits the expression of FOXO1, leading to proliferation, migration, and invasion of breast cancer cells
    Figure Legend Snippet: Proposed mechanistic scheme: miR-29c suppresses breast cancer by the TIMP3/STAT1/FOXO1 pathway. The expression of miR-29c was decreased in breast cancer, which leading to the elevated expression of DNMT3B that is critical for promoter methylation and decreases the expression of TIMP3. As a result, the expression of STAT1 was increased, and it inhibits the expression of FOXO1, leading to proliferation, migration, and invasion of breast cancer cells

    Techniques Used: Expressing, Methylation, Migration

    6) Product Images from "Soluble fibrinogen‐like protein 2 ameliorates acute rejection of liver transplantation in rat via inducing Kupffer cells M2 polarization, et al. Soluble fibrinogen‐like protein 2 ameliorates acute rejection of liver transplantation in rat via inducing Kupffer cells M2 polarization"

    Article Title: Soluble fibrinogen‐like protein 2 ameliorates acute rejection of liver transplantation in rat via inducing Kupffer cells M2 polarization, et al. Soluble fibrinogen‐like protein 2 ameliorates acute rejection of liver transplantation in rat via inducing Kupffer cells M2 polarization

    Journal: Cancer Medicine

    doi: 10.1002/cam4.1528

    sFGL2 induces KCs M2 polarization and inhibits STAT1 and NF‐κB signaling pathway. A, The supernatant levels of IL‐12, TNF‐α, IL‐10, and TGF‐β in control group and r‐FGL2‐treated group were examined by ELISA. B, The frequencies of F4/80 + CD206 + M2 KCs in control and r‐FGL2‐treated groups were detected by flow cytometry. C, The protein levels of STAT1 and p‐STAT1 in control and r‐FGL2‐treated groups were measured using Western blot. D, The protein levels of IκBα, p‐IκBα, NF‐κB p65, p‐p65 in control and r‐FGL2‐treated groups were measured using Western blot. ** P
    Figure Legend Snippet: sFGL2 induces KCs M2 polarization and inhibits STAT1 and NF‐κB signaling pathway. A, The supernatant levels of IL‐12, TNF‐α, IL‐10, and TGF‐β in control group and r‐FGL2‐treated group were examined by ELISA. B, The frequencies of F4/80 + CD206 + M2 KCs in control and r‐FGL2‐treated groups were detected by flow cytometry. C, The protein levels of STAT1 and p‐STAT1 in control and r‐FGL2‐treated groups were measured using Western blot. D, The protein levels of IκBα, p‐IκBα, NF‐κB p65, p‐p65 in control and r‐FGL2‐treated groups were measured using Western blot. ** P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Western Blot

    7) Product Images from "miR-29c plays a suppressive role in breast cancer by targeting the TIMP3/STAT1/FOXO1 pathway"

    Article Title: miR-29c plays a suppressive role in breast cancer by targeting the TIMP3/STAT1/FOXO1 pathway

    Journal: Clinical Epigenetics

    doi: 10.1186/s13148-018-0495-y

    miR-29c directly targeted DNMT3B and regulated the DNMT3B/TIMP3/STAT1/FOXO1 pathway in breast cancer cells. a The potential binding site of miR-29c in the 3′UTR of DNMT3B. b Dual-Luciferase Reporter Assay of miR-29c and DNMT3B in MDA-MB-231 cells. c Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MCF-7 cells after the transfection of miR-29c inhibitor. d qMS-PCR assay of the methylation level of TIMP3 in MCF-7 cells after the transfection of miR-29c inhibitor. e Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MDA-MB-231 cells after the transfection of miR-29c mimic. f qMS-PCR assay of the methylation level of TIMP3 in MDA-MB-231 cells after the transfection of miR-29c mimic. g Migration assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. h Invasion assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. i Protein levels of STAT1 and FOXO1 detected by Western blotting in MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. Data are presented as mean ± SD from three independent experiments, and every experiment was repeated three times, * P
    Figure Legend Snippet: miR-29c directly targeted DNMT3B and regulated the DNMT3B/TIMP3/STAT1/FOXO1 pathway in breast cancer cells. a The potential binding site of miR-29c in the 3′UTR of DNMT3B. b Dual-Luciferase Reporter Assay of miR-29c and DNMT3B in MDA-MB-231 cells. c Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MCF-7 cells after the transfection of miR-29c inhibitor. d qMS-PCR assay of the methylation level of TIMP3 in MCF-7 cells after the transfection of miR-29c inhibitor. e Protein levels of DNMT3B, TIMP3, STAT1, and FOXO1 detected by Western blotting in MDA-MB-231 cells after the transfection of miR-29c mimic. f qMS-PCR assay of the methylation level of TIMP3 in MDA-MB-231 cells after the transfection of miR-29c mimic. g Migration assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. h Invasion assays of MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. i Protein levels of STAT1 and FOXO1 detected by Western blotting in MCF-7 cells that were co-treated with fludarabine (STAT1 inhibitor) and miR-29c inhibitor. Data are presented as mean ± SD from three independent experiments, and every experiment was repeated three times, * P

    Techniques Used: Binding Assay, Luciferase, Reporter Assay, Multiple Displacement Amplification, Western Blot, Transfection, Polymerase Chain Reaction, Methylation, Migration

    Proposed mechanistic scheme: miR-29c suppresses breast cancer by the TIMP3/STAT1/FOXO1 pathway. The expression of miR-29c was decreased in breast cancer, which leading to the elevated expression of DNMT3B that is critical for promoter methylation and decreases the expression of TIMP3. As a result, the expression of STAT1 was increased, and it inhibits the expression of FOXO1, leading to proliferation, migration, and invasion of breast cancer cells
    Figure Legend Snippet: Proposed mechanistic scheme: miR-29c suppresses breast cancer by the TIMP3/STAT1/FOXO1 pathway. The expression of miR-29c was decreased in breast cancer, which leading to the elevated expression of DNMT3B that is critical for promoter methylation and decreases the expression of TIMP3. As a result, the expression of STAT1 was increased, and it inhibits the expression of FOXO1, leading to proliferation, migration, and invasion of breast cancer cells

    Techniques Used: Expressing, Methylation, Migration

    8) Product Images from "CCR2 Signaling Selectively Regulates IFN-α: Role of β-Arrestin 2 in IFNAR1 Internalization"

    Article Title: CCR2 Signaling Selectively Regulates IFN-α: Role of β-Arrestin 2 in IFNAR1 Internalization

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1800598

    β-Arrestin 2 Promotes IFNAR1 Internalization and Inhibits STAT1 Activation.
    Figure Legend Snippet: β-Arrestin 2 Promotes IFNAR1 Internalization and Inhibits STAT1 Activation.

    Techniques Used: Activation Assay

    9) Product Images from "Characterization and allergic role of IL-33-induced neutrophil polarization"

    Article Title: Characterization and allergic role of IL-33-induced neutrophil polarization

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/cmi.2017.163

    The specific phenotype of IL-33-treated neutrophils is mediated by JNK and NF-κB signaling pathways. ( a ) Neutrophils freshly isolated from bone marrow were treated with IL-33 for 15, 30 and 120 min, respectively. The activities of JNK, p38, STAT1, Erk and p65 in cytoplasm were detected by western blotting. ( b ) The c-jun and p65 protein levels in nucleus of neutrophils after IL-33 treatment for 1 and 2 h, respectively, were determined by western blotting. ( c and d ) Neutrophils were pretreated with the indicated concentrations of JNK inhibitor (SP600125) and NF-κB inhibitor for 30 min and then stimulated with IL-33 for 24 h. The levels of IL-9, CCL2, CCL7, CCL12, IL-1R2 and IL-13Ra1 expression were detected by real-time PCR. ( e and f ) The concentrations of IL-9, IL-4, IL-5, IL-13, CCL2 and CCL7 in the supernatant of neutrophils pretreated with either SP600125 or NF-κB inhibitor and then cultured with IL-33 for 24 h were determined by ELISA assay kits. Assays were performed more than three times. Data are shown as mean±s.d. ( n =3). * P
    Figure Legend Snippet: The specific phenotype of IL-33-treated neutrophils is mediated by JNK and NF-κB signaling pathways. ( a ) Neutrophils freshly isolated from bone marrow were treated with IL-33 for 15, 30 and 120 min, respectively. The activities of JNK, p38, STAT1, Erk and p65 in cytoplasm were detected by western blotting. ( b ) The c-jun and p65 protein levels in nucleus of neutrophils after IL-33 treatment for 1 and 2 h, respectively, were determined by western blotting. ( c and d ) Neutrophils were pretreated with the indicated concentrations of JNK inhibitor (SP600125) and NF-κB inhibitor for 30 min and then stimulated with IL-33 for 24 h. The levels of IL-9, CCL2, CCL7, CCL12, IL-1R2 and IL-13Ra1 expression were detected by real-time PCR. ( e and f ) The concentrations of IL-9, IL-4, IL-5, IL-13, CCL2 and CCL7 in the supernatant of neutrophils pretreated with either SP600125 or NF-κB inhibitor and then cultured with IL-33 for 24 h were determined by ELISA assay kits. Assays were performed more than three times. Data are shown as mean±s.d. ( n =3). * P

    Techniques Used: Isolation, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    10) Product Images from "Blimp-1-Mediated Pathway Promotes Type I IFN Production in Plasmacytoid Dendritic Cells by Targeting to Interleukin-1 Receptor-Associated Kinase M"

    Article Title: Blimp-1-Mediated Pathway Promotes Type I IFN Production in Plasmacytoid Dendritic Cells by Targeting to Interleukin-1 Receptor-Associated Kinase M

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01828

    Impaired IKKα and IRF7 activation in plasmacytoid dendritic cells (pDCs) with reduced Blimp-1 expression. (A) Immunoblot analysis of nuclear extracts showing the levels of IRF7 in 4-OHT treated Ctrl-ER and CKO-ER FLpDCs following stimulation with 1 µM CpG-A at indicated time points. (B) Immunoblot analysis of total cell lysates showing the levels of IRF7 phosphorylation at Ser471/472 in 1 µM CpG-A stimulated Ctrl-ER and CKO-ER FLpDCs. (C) Immunoblot analysis showing the levels of phospho-IKKα at Ser176/180, OPN and phospho-AKT at Ser473, in 4-OHT treated and CpG-A stimulated Ctrl-ER and CKO-ER FLpDCs. The quantification of pIKKα was presented by the ratios of pIKKα band intensity vs. IKKα band intensity at each time point. (D) Immunoblot analysis using nuclear extracts of 4-OHT treated Ctrl-ER and CKO-ER FLpDCs showing the levels of p50 and p65 translocation after 1 µM CpG-A treatment at indicated time points. (E) Immunoblot analysis showing the levels of phosphorylated STAT1 at Tyr701 at the indicated time points in Ctrl-ER and CKO-ER FLpDCs treated with 4-OHT and then stimulated with 1 µM CpG-A at the indicated time points. (F) Immunoblot analysis showing levels of phosphorylated STAT1 at Tyr701 in FLpDCs in the presence of 500 U/ml mIFN-α.
    Figure Legend Snippet: Impaired IKKα and IRF7 activation in plasmacytoid dendritic cells (pDCs) with reduced Blimp-1 expression. (A) Immunoblot analysis of nuclear extracts showing the levels of IRF7 in 4-OHT treated Ctrl-ER and CKO-ER FLpDCs following stimulation with 1 µM CpG-A at indicated time points. (B) Immunoblot analysis of total cell lysates showing the levels of IRF7 phosphorylation at Ser471/472 in 1 µM CpG-A stimulated Ctrl-ER and CKO-ER FLpDCs. (C) Immunoblot analysis showing the levels of phospho-IKKα at Ser176/180, OPN and phospho-AKT at Ser473, in 4-OHT treated and CpG-A stimulated Ctrl-ER and CKO-ER FLpDCs. The quantification of pIKKα was presented by the ratios of pIKKα band intensity vs. IKKα band intensity at each time point. (D) Immunoblot analysis using nuclear extracts of 4-OHT treated Ctrl-ER and CKO-ER FLpDCs showing the levels of p50 and p65 translocation after 1 µM CpG-A treatment at indicated time points. (E) Immunoblot analysis showing the levels of phosphorylated STAT1 at Tyr701 at the indicated time points in Ctrl-ER and CKO-ER FLpDCs treated with 4-OHT and then stimulated with 1 µM CpG-A at the indicated time points. (F) Immunoblot analysis showing levels of phosphorylated STAT1 at Tyr701 in FLpDCs in the presence of 500 U/ml mIFN-α.

    Techniques Used: Activation Assay, Expressing, Translocation Assay

    11) Product Images from "The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse"

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI200316603

    The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P
    Figure Legend Snippet: The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P

    Techniques Used:

    Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.
    Figure Legend Snippet: Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.

    Techniques Used: Mouse Assay

    Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.
    Figure Legend Snippet: Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.

    Techniques Used: Mouse Assay, Staining

    Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.
    Figure Legend Snippet: Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.

    Techniques Used: Activity Assay, Mouse Assay, Release Assay

    Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.
    Figure Legend Snippet: Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.

    Techniques Used: Mouse Assay

    Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.
    Figure Legend Snippet: Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.

    Techniques Used: Functional Assay, Expressing, Fluorescence, Staining, Polymerase Chain Reaction, Isolation, Positive Control

    Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).
    Figure Legend Snippet: Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).

    Techniques Used: Mouse Assay, Injection, Mass Spectrometry

    Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.
    Figure Legend Snippet: Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.

    Techniques Used: Staining, Immunohistochemistry, Activation Assay, Flow Cytometry, Expressing, Northern Blot, Polymerase Chain Reaction, Positive Control, Negative Control

    12) Product Images from "The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse"

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI200316603

    The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P
    Figure Legend Snippet: The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P

    Techniques Used:

    Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.
    Figure Legend Snippet: Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.

    Techniques Used: Mouse Assay

    Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.
    Figure Legend Snippet: Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.

    Techniques Used: Mouse Assay, Staining

    Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.
    Figure Legend Snippet: Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.

    Techniques Used: Activity Assay, Mouse Assay, Release Assay

    Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.
    Figure Legend Snippet: Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.

    Techniques Used: Mouse Assay

    Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.
    Figure Legend Snippet: Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.

    Techniques Used: Functional Assay, Expressing, Fluorescence, Staining, Polymerase Chain Reaction, Isolation, Positive Control

    Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).
    Figure Legend Snippet: Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).

    Techniques Used: Mouse Assay, Injection, Mass Spectrometry

    Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.
    Figure Legend Snippet: Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.

    Techniques Used: Staining, Immunohistochemistry, Activation Assay, Flow Cytometry, Expressing, Northern Blot, Polymerase Chain Reaction, Positive Control, Negative Control

    13) Product Images from "The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse"

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse

    Journal: Journal of Clinical Investigation

    doi: 10.1172/JCI200316603

    The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P
    Figure Legend Snippet: The antitumor effects of IFN-α are not enhanced after reconstitution of STAT1 signaling within the melanoma cell. ( a ) IFN-α treatment (open triangles) significantly ( P

    Techniques Used:

    Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.
    Figure Legend Snippet: Effects of IFN-α on the survival of C57BL/6 and STAT1-deficient mice bearing locoregional B16F10 metastases. IFN-α treatment (open triangles) significantly prolonged the survival of ( a ) C57BL/6 mice ( P = 0.002) but not ( b ) STAT1-deficient mice ( P = 0.73) bearing B16F10 locoregional metastases as compared with PBS-treated (filled circles) controls. There were 10 mice in each group.

    Techniques Used: Mouse Assay

    Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.
    Figure Legend Snippet: Histochemical analysis. B16F1 tumors from C57BL/6 mice treated with ( a ) PBS or ( b ) IFN-α and STAT1-deficient mice treated with ( c ) PBS or ( d ) IFN-α were stained with hematoxylin and eosin and analyzed at ×40. A striking degree of tumor cell necrosis was observed in STAT1-deficient mice treated with either PBS or IFN-α. A similar degree of necrosis was observed in PBS-treated C57BL/6 mice. Notably, the least degree of tumor cell necrosis was observed in C57BL/6 mice treated with IFN-α, which was the only group exhibiting prolonged survival.

    Techniques Used: Mouse Assay, Staining

    Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.
    Figure Legend Snippet: Deficient splenocyte cytotoxic activity in STAT1 –/– mice. ( a ) Unstimulated splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the NK-sensitive YAC-1 cell line in a standard 4-hour 51 Cr release assay. ( b ) Splenocytes from STAT1 –/– mice (open circles) and wild-type C57BL/6 mice (filled circles) were tested for cytotoxic activity against the B16F1 murine melanoma cell line in a standard 4-hour 51 Cr release assay after stimulation with 10 4 U/ml IFN-α for 18 hours.

    Techniques Used: Activity Assay, Mouse Assay, Release Assay

    Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.
    Figure Legend Snippet: Survival of mice bearing the STAT1-deficient AGS-1 murine melanoma cell line. IFN-α treatment (open triangles) prolonged the survival of C57BL/6 mice ( n = 6 mice per group) challenged with the STAT1-deficient AGS-1 melanoma cell line as compared with PBS-treated mice (filled circles) ( P = 0.008). These results are representative of two separate determinations.

    Techniques Used: Mouse Assay

    Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.
    Figure Legend Snippet: Reconstitution of functional STAT1 in the AGS-1 cell line. ( a ) Immunoblot analysis of lysates from AGS-1 STAT1 cells using an anti-STAT1 antibody indicated that expression of STAT1 protein was restored. Ctrl, control. ( b ) The ability of STAT1 protein from AGS-1 STAT1 cells to undergo phosphorylation at Tyr-701 was confirmed by immunoblotting with an anti-P-STAT1 monoclonal antibody. ( c ) Overnight stimulation with IFN-α (10 3 U/ml) resulted in enhanced H2k expression on AGS-1 STAT1 but not AGS-1 MSCV melanoma cells. Shaded histograms represent fluorescence intensity of H2k-PE staining in IFN-α–stimulated EGFP-positive cells. White histograms represent H2k-PE staining in PBS-stimulated EGFP-positive cells. Determination of H2k-positive cells was based on isotype control antibodies (M1), which fell within the first log of fluorescence. ( d ) IFN-α inhibits proliferation of the AGS-1 STAT1 cell line in a dose-dependent manner. In contrast, the antiproliferative effects of IFN-α were not evident in AGS-1 MSCV cells. Results represent the mean of triplicate wells and are expressed as optical density at 570 nm ± SE. ( e ) The ability of the AGS-1 STAT1 cell line to regulate ISGF3 gene transcription in response to IFN-α was confirmed by PCR. Total cellular RNA was isolated from cell lines (B16F1, AGS-1, AGS-1 STAT1 , and AGS-1 MSCV ) after overnight treatment with 10 4 U/ml of murine IFN-α and converted to cDNA using standard methods. PCR primers specific for muISG-54 detected increased ISG-54 expression in only the B16F1 (positive control) and AGS-1 STAT1 cell lines after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as controls.

    Techniques Used: Functional Assay, Expressing, Fluorescence, Staining, Polymerase Chain Reaction, Isolation, Positive Control

    Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).
    Figure Legend Snippet: Absence of STAT1 in the host results in decreased signaling in response to IFN-α and decreased survival after tumor challenge. ( a ) A DNA probe specific for activated murine STAT1 reacted with whole-cell lysates from IFN-α–treated splenocytes of C57BL/6 but not STAT1-deficient mice. Similar results were obtained in other tissues, including liver and kidney (data not shown). STAT1 –/– and C57BL/6 mice ( n = 10 mice per group) were injected intraperitoneally with 10 6 B16F1 melanoma cells. As compared with PBS-treated controls (filled circles), IFN-α treatment (2 × 10 4 U per day) (open triangles) prolonged the survival of C57BL/6 mice but not STAT1 –/– mice challenged with B16F1 melanoma cells ( b and c ) or JB/MS melanoma cells ( n = 6 mice per group) ( d and e ).

    Techniques Used: Mouse Assay, Injection, Mass Spectrometry

    Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.
    Figure Legend Snippet: Characterization of the AGS-1 cell line. AGS-1 has the appearance of a malignant melanoma by ( a ) hematoxylin and eosin staining and immunohistochemistry for ( b ) S-100 and ( c ) HMB-45. ( d ) Immunoblot analysis confirmed that the AGS-1 cell line was STAT1 deficient (lane 3). Cell lysates from the A431 cell line (lane 1) and B16F1 cell line (lane 2) were used as positive controls. The protein content of the lysates was quantitated, and equal amounts were loaded onto the gel. ( e ) EMSA analysis revealed no activation of STAT1 in IFN-α–treated AGS-1 cells. Lysates from IFN-α–stimulated B16F1 cells were used as positive controls in this assay. SIS, SIS-inducible element. ( f ) Flow cytometric analysis of H2k expression. Treatment of B16F1 cells for 48 hours with 10 4 ). ( g ) Northern blot analysis indicated a loss of ISG-54 gene regulation in AGS-1 cells after stimulation with 10 4 U/ml muIFN-α for 16 hours. Probes specific for GAPDH (housekeeping gene) were used as controls and did not vary across samples. ( h ) Northern blot findings were confirmed by PCR using primers specific for muISG-54 and showed increased ISG-54 expression only in B16F1 cells (positive control) after IFN-α treatment. Primers specific for 28s rRNA (housekeeping gene) were used as a controls in this assay. Neg, negative control.

    Techniques Used: Staining, Immunohistochemistry, Activation Assay, Flow Cytometry, Expressing, Northern Blot, Polymerase Chain Reaction, Positive Control, Negative Control

    14) Product Images from "Rit Signaling Contributes to Interferon-γ-Induced Dendritic Retraction via p38 MAP Kinase Activation"

    Article Title: Rit Signaling Contributes to Interferon-γ-Induced Dendritic Retraction via p38 MAP Kinase Activation

    Journal: Journal of neurochemistry

    doi: 10.1111/j.1471-4159.2008.05708.x

    IFNγ-mediated STAT activation does not require p38 signaling On day 7 in vitro , hippocampal neurons were treated with IFNγ (30 ng/ml) for 30 min in the absence or presence of the p38 MAP kinase inhibitor, SB203580 (10 µM). Cell lysates were analyzed by immunoblotting using phosphospecific antibodies for STAT1. A representative Western blot ( top panel ) and corresponding densitometric analysis ( bottom panel ) of STAT1 phosphorylated on Ser 727 (pSTAT) and total (phosphorylated and non-phosphorylated) STAT1 indicate that SB203580 does not block IFNγ activation of STAT1. Data are expressed as mean ± SEM (n=3 blots obtained in 3 separate experiments performed using cultures from 3 separate dissections).
    Figure Legend Snippet: IFNγ-mediated STAT activation does not require p38 signaling On day 7 in vitro , hippocampal neurons were treated with IFNγ (30 ng/ml) for 30 min in the absence or presence of the p38 MAP kinase inhibitor, SB203580 (10 µM). Cell lysates were analyzed by immunoblotting using phosphospecific antibodies for STAT1. A representative Western blot ( top panel ) and corresponding densitometric analysis ( bottom panel ) of STAT1 phosphorylated on Ser 727 (pSTAT) and total (phosphorylated and non-phosphorylated) STAT1 indicate that SB203580 does not block IFNγ activation of STAT1. Data are expressed as mean ± SEM (n=3 blots obtained in 3 separate experiments performed using cultures from 3 separate dissections).

    Techniques Used: Activation Assay, In Vitro, Western Blot, Blocking Assay

    Rit activates p38, but neither ERK1/2 nor STAT signaling (A and B) Hippocampal neurons were infected with adenovirus expressing GFP alone (Ad-GFP) or co-expressing GFP and constitutively active RitQ79L (Ad-caRit) 4 days after plating. Cell lysates were collected 16 h post-infection and analyzed by immunoblotting using mAb that specifically recognize phosphorylated Ser 727 on STAT1 (pSTAT1) or polyclonal antibodies that react with both phosphorylated and nonphosphorylated STAT1 (STAT1). A representative Western blot (A) and corresponding densitometric analysis (B) of pSTAT1 and total STAT1 indicate that treatment with IFNγ but not expression of caRit activates STAT1 as indicated by increased levels of pSTAT1. Data are expressed as mean ± SEM (n=3 blots obtained in 3 separate experiments performed using cultures from 3 separate dissections). (C) IFNγ activates p38 but not ERK1/2 in PC6 cells. PC6 cells were transfected with either shCTR or shRit208 RNAi vectors and subjected to G418 selection for 48 h. Cells were then serum starved (serum-free DMEM, 5 h) prior to stimulation with IFNγ (50 ng/ml). Whole cell lysates were prepared and levels of activated p38 and ERK1/2 determined by phosphospecific immunoblotting. A representative Western blot for activated p38 and ERK1/2 MAP kinase levels is shown. Anti-actin immunoblotting was used to confirm equivalent protein loading. Note that IFNγ fails to stimulate ERK MAP kinase signaling in PC6 cells and that Rit silencing inhibits IFNγ-mediated p38 activation.
    Figure Legend Snippet: Rit activates p38, but neither ERK1/2 nor STAT signaling (A and B) Hippocampal neurons were infected with adenovirus expressing GFP alone (Ad-GFP) or co-expressing GFP and constitutively active RitQ79L (Ad-caRit) 4 days after plating. Cell lysates were collected 16 h post-infection and analyzed by immunoblotting using mAb that specifically recognize phosphorylated Ser 727 on STAT1 (pSTAT1) or polyclonal antibodies that react with both phosphorylated and nonphosphorylated STAT1 (STAT1). A representative Western blot (A) and corresponding densitometric analysis (B) of pSTAT1 and total STAT1 indicate that treatment with IFNγ but not expression of caRit activates STAT1 as indicated by increased levels of pSTAT1. Data are expressed as mean ± SEM (n=3 blots obtained in 3 separate experiments performed using cultures from 3 separate dissections). (C) IFNγ activates p38 but not ERK1/2 in PC6 cells. PC6 cells were transfected with either shCTR or shRit208 RNAi vectors and subjected to G418 selection for 48 h. Cells were then serum starved (serum-free DMEM, 5 h) prior to stimulation with IFNγ (50 ng/ml). Whole cell lysates were prepared and levels of activated p38 and ERK1/2 determined by phosphospecific immunoblotting. A representative Western blot for activated p38 and ERK1/2 MAP kinase levels is shown. Anti-actin immunoblotting was used to confirm equivalent protein loading. Note that IFNγ fails to stimulate ERK MAP kinase signaling in PC6 cells and that Rit silencing inhibits IFNγ-mediated p38 activation.

    Techniques Used: Infection, Expressing, Western Blot, Transfection, Selection, Activation Assay

    15) Product Images from "Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush"

    Article Title: Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2016.10.009

    Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P
    Figure Legend Snippet: Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P

    Techniques Used: Mouse Assay, Isolation, Western Blot, Injection, Expressing, Real-time Polymerase Chain Reaction

    16) Product Images from "Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush"

    Article Title: Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2016.10.009

    Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P
    Figure Legend Snippet: Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P

    Techniques Used: Mouse Assay, Isolation, Western Blot, Injection, Expressing, Real-time Polymerase Chain Reaction

    17) Product Images from "Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush"

    Article Title: Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2016.10.009

    Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P
    Figure Legend Snippet: Traumatic optic neuropathy (TON)-induced CXCL10 up-regulation is modulated by STAT signaling. A: Wild-type (WT) mice were subjected to TON, and proteins were isolated from retinas at indicated time points after TON. The phosphorylation levels of STAT1, STAT2, STAT3, STAT5, and STAT6 were evaluated by Western blotting. α-Tubulin was used as loading control. B and C: STAT1/STAT3 inhibitor Stattic or vehicle (Veh) was injected intravitreally into WT mice at 1 hour after the induction of TON. Retinas were collected at 4 hours after TON. B: Phosphorylated and total STAT1 and STAT3 were assessed by Western blotting. C: The expression of CXCL10 mRNA was analyzed by quantitative PCR. n = 5 mice ( A–C ). ∗ P

    Techniques Used: Mouse Assay, Isolation, Western Blot, Injection, Expressing, Real-time Polymerase Chain Reaction

    18) Product Images from "Genome and Infection Characteristics of Human Parechovirus Type 1: The Interplay between Viral Infection and Type I Interferon Antiviral System"

    Article Title: Genome and Infection Characteristics of Human Parechovirus Type 1: The Interplay between Viral Infection and Type I Interferon Antiviral System

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0116158

    HPeV1 downregulates type I IFN activity. (A) A549 and T84 cells (3×10 5 ) were infected with mock or HPeV1 at MOI = 5 for 6 h, then treated with IFNβ (1000 U/ml) for 18 h; untreated cells are indicated as ctrl. Immunoblotting of levels of phospho-STAT1 and total STAT1 in whole cell extracts. HPeV1 VP0 indicates viral infection and β-actin is a loading control. (B) Interferon-stimulated response element (ISRE) reporter assay was performed in A549 and T84 cells (3×10 5 ) transfected with ISRE-luciferase reporter plasmids (400 ng) and control vector pRL-TK (40 ng) for 24 h, then infected with HPeV1 for 6 h. After 24 h of IFNβ (1000 U/ml) stimulation, ISRE luciferase activity was measured by dual-luciferase assay and (C) RT-qPCR of mRNA expression. Data are mean ± SD of at least 3 independent experiments. Student t test, * p
    Figure Legend Snippet: HPeV1 downregulates type I IFN activity. (A) A549 and T84 cells (3×10 5 ) were infected with mock or HPeV1 at MOI = 5 for 6 h, then treated with IFNβ (1000 U/ml) for 18 h; untreated cells are indicated as ctrl. Immunoblotting of levels of phospho-STAT1 and total STAT1 in whole cell extracts. HPeV1 VP0 indicates viral infection and β-actin is a loading control. (B) Interferon-stimulated response element (ISRE) reporter assay was performed in A549 and T84 cells (3×10 5 ) transfected with ISRE-luciferase reporter plasmids (400 ng) and control vector pRL-TK (40 ng) for 24 h, then infected with HPeV1 for 6 h. After 24 h of IFNβ (1000 U/ml) stimulation, ISRE luciferase activity was measured by dual-luciferase assay and (C) RT-qPCR of mRNA expression. Data are mean ± SD of at least 3 independent experiments. Student t test, * p

    Techniques Used: Activity Assay, Infection, Reporter Assay, Transfection, Luciferase, Plasmid Preparation, Quantitative RT-PCR, Expressing

    19) Product Images from "Glycogen synthase kinase-3 promotes the synergistic action of interferon-? on lipopolysaccharide-induced IL-6 production in RAW264.7 cells1"

    Article Title: Glycogen synthase kinase-3 promotes the synergistic action of interferon-? on lipopolysaccharide-induced IL-6 production in RAW264.7 cells1

    Journal: Cellular signalling

    doi: 10.1016/j.cellsig.2009.02.019

    IFNγ activates STAT1 and STAT3
    Figure Legend Snippet: IFNγ activates STAT1 and STAT3

    Techniques Used:

    20) Product Images from "CNOT7 depletion reverses natural killer cell resistance by modulating the tumor immune microenvironment of hepatocellular carcinoma"

    Article Title: CNOT7 depletion reverses natural killer cell resistance by modulating the tumor immune microenvironment of hepatocellular carcinoma

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12836

    Changes in CNOT7, STAT1, TGF‐β1 and IFN‐γ levels in patients with HCCBC. (A) Correlation of CNOT7 expression level on HCC patient survival. Red lines represent the high expression of CNOT7 , and blue lines represent low expression. (B) Quantitative data (mean ± SD) of ELISAs were shown. Multiple comparisons were made by the Student‐Newman‐Keuls q (SNK‐ q ) method. * P
    Figure Legend Snippet: Changes in CNOT7, STAT1, TGF‐β1 and IFN‐γ levels in patients with HCCBC. (A) Correlation of CNOT7 expression level on HCC patient survival. Red lines represent the high expression of CNOT7 , and blue lines represent low expression. (B) Quantitative data (mean ± SD) of ELISAs were shown. Multiple comparisons were made by the Student‐Newman‐Keuls q (SNK‐ q ) method. * P

    Techniques Used: Expressing

    21) Product Images from "NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease"

    Article Title: NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease

    Journal: Redox Biology

    doi: 10.1016/j.redox.2017.02.016

    H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p
    Figure Legend Snippet: H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p

    Techniques Used: Western Blot

    Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.
    Figure Legend Snippet: Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.

    Techniques Used: Derivative Assay, Activation Assay, Inhibition, Generated

    22) Product Images from "NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease"

    Article Title: NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease

    Journal: Redox Biology

    doi: 10.1016/j.redox.2017.02.016

    H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p
    Figure Legend Snippet: H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p

    Techniques Used: Western Blot

    Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.
    Figure Legend Snippet: Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.

    Techniques Used: Derivative Assay, Activation Assay, Inhibition, Generated

    23) Product Images from "NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease"

    Article Title: NADPH oxidase-derived H2O2 mediates the regulatory effects of microglia on astrogliosis in experimental models of Parkinson's disease

    Journal: Redox Biology

    doi: 10.1016/j.redox.2017.02.016

    H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p
    Figure Legend Snippet: H 2 O 2 regulates the immunological functions of astrogliosis in a STAT1/3-dependent manner. (A) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in H 2 O 2 -treated astroglia cultures at 1 h of treatment by Western blot and the density of the blot was quantified. (B) The levels of phosphorylated and nonphosphorylated STAT1 and 3 were determined in astroglia culture at 30, 60 and 90 mins of H 2 O 2 treatment by Western blot and the density of the blot was quantified. (C) The expressions of GFAP were determined in H 2 O 2 -treated astroglia cultures with or without AG490 pre-treatment by Western blot and the density of the blot was quantified. (D) The gene expressions of TNFα, iNOS, GDNF and BDNF were determined H 2 O 2 -treated astroglia cultures with or without AG490. Results are expressed as the percentage of controls from three experiments performed in duplicate. * p

    Techniques Used: Western Blot

    Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.
    Figure Legend Snippet: Proposed mechanism showing how microglial NOX2-derived H 2 O 2 regulates astrogliosis. Microglial activation induced by LPS or MPTP precedes astrogliosis. NOX2, a key superoxide-producing enzyme in microglia, is previously reported to play a pivotal role in the initiation of microglial activation leading to neuroinflammation. Pharmacological inhibition or genetic deletion of NOX2 attenuates microglial activation, which is associated with suppression of astrogliosis. NOX2-derived H 2 O 2 directly activates astroglia via a process in which extracellularly generated H 2 O 2 diffuses into the cytoplasm and subsequently enhances phosphorylation of transcription factor, STAT1 and STAT3. Activation of STATs regulates of the gene expressions of both pro-inflammatory and neurotropic factors, therefore, regulating the immunological functions of astrogliosis.

    Techniques Used: Derivative Assay, Activation Assay, Inhibition, Generated

    24) Product Images from "STAT1-mediated inhibition of FOXM1 enhances gemcitabine sensitivity in pancreatic cancer"

    Article Title: STAT1-mediated inhibition of FOXM1 enhances gemcitabine sensitivity in pancreatic cancer

    Journal: Clinical Science (London, England : 1979)

    doi: 10.1042/CS20180816

    IFNγ inhibits FOXM1 sensitization to gemcitabine in pancreatic xenograft tumors ( A,B ) SW1990 cells were subcutaneously injected into the left flank of nude mice. Administration of chemotherapy began when the tumor diameter reached 3–5 mm. The mice were randomly divided into four groups ( n =5) and treated as described in figure. (A) Tumor size was measured after approximately 30 days of treatment. (B) Tumor volumes were measured every 3 days. A tumor growth curve was drawn according the measured tumor volume. ( C ) The representative tumor tissue sections from xenografts in different treatment groups, analyzed by IHC for the expression of FOXM1, pSTAT1, the proliferation markers PCNA and apoptotic marker Cleaved Caspase3. Scale bar, 20 μm. ( D ) A schematic model showing the potential roles of IFNγ in enhancing the sensitivity of pancreatic cancer cells to gemcitabine. IFNγ suppresses the expression of FOXM1 by activating STAT1, causing inhibition of NFκB signaling, which results in increased sensitivity of pancreatic cancer cells to gemcitabine. ** P
    Figure Legend Snippet: IFNγ inhibits FOXM1 sensitization to gemcitabine in pancreatic xenograft tumors ( A,B ) SW1990 cells were subcutaneously injected into the left flank of nude mice. Administration of chemotherapy began when the tumor diameter reached 3–5 mm. The mice were randomly divided into four groups ( n =5) and treated as described in figure. (A) Tumor size was measured after approximately 30 days of treatment. (B) Tumor volumes were measured every 3 days. A tumor growth curve was drawn according the measured tumor volume. ( C ) The representative tumor tissue sections from xenografts in different treatment groups, analyzed by IHC for the expression of FOXM1, pSTAT1, the proliferation markers PCNA and apoptotic marker Cleaved Caspase3. Scale bar, 20 μm. ( D ) A schematic model showing the potential roles of IFNγ in enhancing the sensitivity of pancreatic cancer cells to gemcitabine. IFNγ suppresses the expression of FOXM1 by activating STAT1, causing inhibition of NFκB signaling, which results in increased sensitivity of pancreatic cancer cells to gemcitabine. ** P

    Techniques Used: Injection, Mouse Assay, Immunohistochemistry, Expressing, Marker, Inhibition

    STAT1 directly suppresses FOXM1 expression in pancreatic cancer cells ( A ) Staining of the same cohorts of pancreatic tumor sections for FOXM1 expression and pSTAT1 level. ( B ) The negative correlation of FOXM1 expression with the pSTAT1 level, as assessed using Pearson correlation coefficient analysis ( n =37, r = −0.417, P
    Figure Legend Snippet: STAT1 directly suppresses FOXM1 expression in pancreatic cancer cells ( A ) Staining of the same cohorts of pancreatic tumor sections for FOXM1 expression and pSTAT1 level. ( B ) The negative correlation of FOXM1 expression with the pSTAT1 level, as assessed using Pearson correlation coefficient analysis ( n =37, r = −0.417, P

    Techniques Used: Expressing, Staining

    25) Product Images from "Multiple tumor suppressors regulate a HIF-dependent negative feedback loop via ISGF3 in human clear cell renal cancer"

    Article Title: Multiple tumor suppressors regulate a HIF-dependent negative feedback loop via ISGF3 in human clear cell renal cancer

    Journal: eLife

    doi: 10.7554/eLife.37925

    The correlations between ISGF3 subunits and PBRM1, KDM5C, BAP1 or SEDT2 in the TCGA dataset. Correlations between IRF9, STAT1 or STAT2 with indicated genes within the TCGA dataset. N = 533.
    Figure Legend Snippet: The correlations between ISGF3 subunits and PBRM1, KDM5C, BAP1 or SEDT2 in the TCGA dataset. Correlations between IRF9, STAT1 or STAT2 with indicated genes within the TCGA dataset. N = 533.

    Techniques Used:

    26) Product Images from "ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression"

    Article Title: ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-11-1262

    COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers
    Figure Legend Snippet: COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers

    Techniques Used:

    ANGPTL4 enhances cancer cell growth through STAT1
    Figure Legend Snippet: ANGPTL4 enhances cancer cell growth through STAT1

    Techniques Used:

    27) Product Images from "ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression"

    Article Title: ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-11-1262

    COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers
    Figure Legend Snippet: COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers

    Techniques Used:

    ANGPTL4 enhances cancer cell growth through STAT1
    Figure Legend Snippet: ANGPTL4 enhances cancer cell growth through STAT1

    Techniques Used:

    28) Product Images from "ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression"

    Article Title: ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-11-1262

    COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers
    Figure Legend Snippet: COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers

    Techniques Used:

    ANGPTL4 enhances cancer cell growth through STAT1
    Figure Legend Snippet: ANGPTL4 enhances cancer cell growth through STAT1

    Techniques Used:

    29) Product Images from "ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression"

    Article Title: ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-11-1262

    COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers
    Figure Legend Snippet: COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers

    Techniques Used:

    ANGPTL4 enhances cancer cell growth through STAT1
    Figure Legend Snippet: ANGPTL4 enhances cancer cell growth through STAT1

    Techniques Used:

    30) Product Images from "ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression"

    Article Title: ANGPTL4 induction by prostaglandin E2 under hypoxic conditions promotes colorectal cancer progression

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-11-1262

    COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers
    Figure Legend Snippet: COX-2, ANGPTL4 and STAT1 expressions in colorectal cancers

    Techniques Used:

    ANGPTL4 enhances cancer cell growth through STAT1
    Figure Legend Snippet: ANGPTL4 enhances cancer cell growth through STAT1

    Techniques Used:

    31) Product Images from "MicroRNA-513 Regulates B7-H1 Translation and Is Involved in IFN-γ-Induced B7-H1 Expression in Cholangiocytes 1-Induced B7-H1 Expression in Cholangiocytes 1 , 2"

    Article Title: MicroRNA-513 Regulates B7-H1 Translation and Is Involved in IFN-γ-Induced B7-H1 Expression in Cholangiocytes 1-Induced B7-H1 Expression in Cholangiocytes 1 , 2

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    IFN- γ alters miRNA expression profile in cholangiocytes and decreases miR-513 expression in cholangiocytes in a STAT1-dependent manner. A , Microarray analysis of miRNA expression in H69 cells after IFN- γ stimulation for 8 h. Data were
    Figure Legend Snippet: IFN- γ alters miRNA expression profile in cholangiocytes and decreases miR-513 expression in cholangiocytes in a STAT1-dependent manner. A , Microarray analysis of miRNA expression in H69 cells after IFN- γ stimulation for 8 h. Data were

    Techniques Used: Expressing, Microarray

    IFN-γ alters miRNA expression profile in cholangiocytes and decreases miR-513 expression in a STAT1-dependent manner
    Figure Legend Snippet: IFN-γ alters miRNA expression profile in cholangiocytes and decreases miR-513 expression in a STAT1-dependent manner

    Techniques Used: Expressing

    32) Product Images from "Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿"

    Article Title: Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02610-08

    Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.
    Figure Legend Snippet: Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.

    Techniques Used: Mutagenesis, Transfection, Immunoprecipitation

    Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described
    Figure Legend Snippet: Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described

    Techniques Used: Inhibition, Binding Assay, Construct, Activity Assay

    Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,
    Figure Legend Snippet: Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,

    Techniques Used: Mutagenesis, Infection

    WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at
    Figure Legend Snippet: WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at

    Techniques Used: Mutagenesis, Infection, Transfection, Plasmid Preparation

    NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody
    Figure Legend Snippet: NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody

    Techniques Used: Transfection, Construct, Immunoprecipitation

    33) Product Images from "Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿"

    Article Title: Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02610-08

    Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.
    Figure Legend Snippet: Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.

    Techniques Used: Mutagenesis, Transfection, Immunoprecipitation

    Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described
    Figure Legend Snippet: Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described

    Techniques Used: Inhibition, Binding Assay, Construct, Activity Assay

    Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,
    Figure Legend Snippet: Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,

    Techniques Used: Mutagenesis, Infection

    WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at
    Figure Legend Snippet: WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at

    Techniques Used: Mutagenesis, Infection, Transfection, Plasmid Preparation

    NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody
    Figure Legend Snippet: NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody

    Techniques Used: Transfection, Construct, Immunoprecipitation

    34) Product Images from "Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿"

    Article Title: Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02610-08

    Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.
    Figure Legend Snippet: Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.

    Techniques Used: Mutagenesis, Transfection, Immunoprecipitation

    Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described
    Figure Legend Snippet: Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described

    Techniques Used: Inhibition, Binding Assay, Construct, Activity Assay

    Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,
    Figure Legend Snippet: Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,

    Techniques Used: Mutagenesis, Infection

    WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at
    Figure Legend Snippet: WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at

    Techniques Used: Mutagenesis, Infection, Transfection, Plasmid Preparation

    NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody
    Figure Legend Snippet: NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody

    Techniques Used: Transfection, Construct, Immunoprecipitation

    35) Product Images from "Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿"

    Article Title: Nipah Virus Sequesters Inactive STAT1 in the Nucleus via a P Gene-Encoded Mechanism ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.02610-08

    Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.
    Figure Legend Snippet: Point mutant forms of NiV V and W do not interact with or inhibit STAT1. (A) Plasmids encoding the HA-tagged NiV V and W WT or mutant proteins were transfected in 293T cells and immunoprecipitated (IP) as described in the legend to Fig.

    Techniques Used: Mutagenesis, Transfection, Immunoprecipitation

    Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described
    Figure Legend Snippet: Single amino acid substitutions decrease NiV P's inhibition of IFN signaling and STAT1 binding. HA NiV P constructs harboring the indicated point mutations were assayed for polymerase cofactor activity, IFN signaling inhibition, and STAT1 binding as described

    Techniques Used: Inhibition, Binding Assay, Construct, Activity Assay

    Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,
    Figure Legend Snippet: Impact of WT and mutant NiVs on the localization and phosphorylation of endogenous STAT1. Vero E6 cells were infected with the indicated viruses and treated with IFN-β at 12 h postinfection. The phospho-Y701 form of endogenous STAT1 (P-STAT1,

    Techniques Used: Mutagenesis, Infection

    WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at
    Figure Legend Snippet: WT and C ko viruses direct STAT1 to the nucleus, but the G121E mutant virus fails to control STAT1 localization. STAT1-GFP localization following NiV infection and subsequent IFN treatment. Vero E6 cells were transfected with STAT1-GFP plasmid, and at

    Techniques Used: Mutagenesis, Infection, Transfection, Plasmid Preparation

    NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody
    Figure Legend Snippet: NiV P, V, and W IFN signaling mutants fail to bind and inhibit STAT1. (A) 293T cells were transfected with HA-tagged NiV P WT or internal deletion constructs spanning residues 51 to 150. At 24 hpt, immunoprecipitation (IP) was performed with anti-HA antibody

    Techniques Used: Transfection, Construct, Immunoprecipitation

    36) Product Images from "Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages"

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.286427

    STAT1 is required for IFNγ-stimulated IRF8 expression. IRF8 mRNA was detected in RAW264.7 cells after transfection with IRF8 shRNA (shIRF8–1 and shIRF8–2) or STAT1 shRNA plasmids (shSTAT1). As a control, the cells were transfected
    Figure Legend Snippet: STAT1 is required for IFNγ-stimulated IRF8 expression. IRF8 mRNA was detected in RAW264.7 cells after transfection with IRF8 shRNA (shIRF8–1 and shIRF8–2) or STAT1 shRNA plasmids (shSTAT1). As a control, the cells were transfected

    Techniques Used: Expressing, Transfection, shRNA

    Roles of PU.1, IRF8, STAT1, and IRF1 in IFNγ stimulation of the C1qB promoter. RAW264.7 cells were transfected with the B273 promoter and also co-transfected with plasmids encoding shRNA for mouse IRF1 ( A ), STAT1 ( B ), IRF8 (IRF8–1, IRF8–2;
    Figure Legend Snippet: Roles of PU.1, IRF8, STAT1, and IRF1 in IFNγ stimulation of the C1qB promoter. RAW264.7 cells were transfected with the B273 promoter and also co-transfected with plasmids encoding shRNA for mouse IRF1 ( A ), STAT1 ( B ), IRF8 (IRF8–1, IRF8–2;

    Techniques Used: Transfection, shRNA

    37) Product Images from "Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages"

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.286427

    STAT1 is required for IFNγ-stimulated IRF8 expression. IRF8 mRNA was detected in RAW264.7 cells after transfection with IRF8 shRNA (shIRF8–1 and shIRF8–2) or STAT1 shRNA plasmids (shSTAT1). As a control, the cells were transfected
    Figure Legend Snippet: STAT1 is required for IFNγ-stimulated IRF8 expression. IRF8 mRNA was detected in RAW264.7 cells after transfection with IRF8 shRNA (shIRF8–1 and shIRF8–2) or STAT1 shRNA plasmids (shSTAT1). As a control, the cells were transfected

    Techniques Used: Expressing, Transfection, shRNA

    Roles of PU.1, IRF8, STAT1, and IRF1 in IFNγ stimulation of the C1qB promoter. RAW264.7 cells were transfected with the B273 promoter and also co-transfected with plasmids encoding shRNA for mouse IRF1 ( A ), STAT1 ( B ), IRF8 (IRF8–1, IRF8–2;
    Figure Legend Snippet: Roles of PU.1, IRF8, STAT1, and IRF1 in IFNγ stimulation of the C1qB promoter. RAW264.7 cells were transfected with the B273 promoter and also co-transfected with plasmids encoding shRNA for mouse IRF1 ( A ), STAT1 ( B ), IRF8 (IRF8–1, IRF8–2;

    Techniques Used: Transfection, shRNA

    38) Product Images from "The breast cancer oncogene IKKε coordinates mitochondrial function and serine metabolism"

    Article Title: The breast cancer oncogene IKKε coordinates mitochondrial function and serine metabolism

    Journal: EMBO reports

    doi: 10.15252/embr.201948260

    IKKε mediates cytokine secretion but no autocrine effect is responsible for the induction of SBP enzymes (A-C) Representative western blots showing levels of IKKε, PHGDH, PSAT1, PSPH, STAT1, phosphorylated STAT1 (Y701), OAS1, p65 and phosphorylated p65 (S486) in (A) Flp-In 293 HA-GFP cells, (B) T47D breast cancer cells and (C) ZR-75-1 breast cancer cells treated for 24 hours with media conditioned by Flp-In 293 HA-GFP or Flp-In 293 HA-IKKε cells treated with doxycycline (Dox) for 16 hours. Changes in protein expression were compared to changes in Flp-In 293 HA-IKKε cells treated with doxycycline (16 hours).
    Figure Legend Snippet: IKKε mediates cytokine secretion but no autocrine effect is responsible for the induction of SBP enzymes (A-C) Representative western blots showing levels of IKKε, PHGDH, PSAT1, PSPH, STAT1, phosphorylated STAT1 (Y701), OAS1, p65 and phosphorylated p65 (S486) in (A) Flp-In 293 HA-GFP cells, (B) T47D breast cancer cells and (C) ZR-75-1 breast cancer cells treated for 24 hours with media conditioned by Flp-In 293 HA-GFP or Flp-In 293 HA-IKKε cells treated with doxycycline (Dox) for 16 hours. Changes in protein expression were compared to changes in Flp-In 293 HA-IKKε cells treated with doxycycline (16 hours).

    Techniques Used: Western Blot, Expressing

    39) Product Images from "p16INK4a deficiency promotes IL-4-induced polarization and inhibits proinflammatory signaling in macrophages"

    Article Title: p16INK4a deficiency promotes IL-4-induced polarization and inhibits proinflammatory signaling in macrophages

    Journal: Blood

    doi: 10.1182/blood-2010-10-313106

    p16 INK4a -deficiency results in an alteration of STAT1 and NF-κB signaling Representation of the relative microarray intensity values from a selection of down-regulated genes in p16 +/+ and p16 −/− BMDM with or without polarization (AAMφ) by 15 ng/mL IL-4 from day 0 of differentiation. Statistically significant differences are indicated (a: p
    Figure Legend Snippet: p16 INK4a -deficiency results in an alteration of STAT1 and NF-κB signaling Representation of the relative microarray intensity values from a selection of down-regulated genes in p16 +/+ and p16 −/− BMDM with or without polarization (AAMφ) by 15 ng/mL IL-4 from day 0 of differentiation. Statistically significant differences are indicated (a: p

    Techniques Used: Microarray, Selection

    40) Product Images from "Pellino1 Is Required for Interferon Production by Viral Double-stranded RNA *"

    Article Title: Pellino1 Is Required for Interferon Production by Viral Double-stranded RNA *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M112.367557

    The IFNβ-stimulated production of IFNβ-regulated genes is suppressed in BMDM from Pellino1[F397A] mice “downstream” of the activation of the JAK-STAT1/2 pathway. A , BMDM from wild type ( WT ) or Pellino1[F397A] mice were
    Figure Legend Snippet: The IFNβ-stimulated production of IFNβ-regulated genes is suppressed in BMDM from Pellino1[F397A] mice “downstream” of the activation of the JAK-STAT1/2 pathway. A , BMDM from wild type ( WT ) or Pellino1[F397A] mice were

    Techniques Used: Mouse Assay, Activation Assay

    Related Articles

    Mouse Assay:

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse
    Article Snippet: .. Identical results were obtained in a second murine model of malignant melanoma in which a highly metastatic melanoma cell line (B16F10) was used to generate locoregional metastases in the inguinal lymph nodes of STAT1 –/– mice. .. STAT1 –/– mice exhibited normal levels of circulating immune effector cells, but splenocytes from STAT1 –/– mice exhibited a reduction in cytotoxic activity against the B16 melanoma cell line and the NK-sensitive YAC-1 cell line.

    Activation Assay:

    Article Title: Genome and Infection Characteristics of Human Parechovirus Type 1: The Interplay between Viral Infection and Type I Interferon Antiviral System
    Article Snippet: .. Previous study revealed enterovirus 71 2A protease reduces IFN receptor 1 activity to disrupt the activation of STAT1, STAT2, Jak1 and Tyk2 [ ]. .. These data may suggest future investigation of HPeV1 regulating type I IFN activity because we also found the HPeV1 can attenuate type I IFN downstream signaling and antiviral protein expression.

    other:

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse
    Article Snippet: The presence of STAT1 was confirmed in AGS-1STAT1 cells by immunoblot analysis (Figure a).

    Article Title: The antitumor effects of IFN-? are abrogated in a STAT1-deficient mouse
    Article Snippet: Interestingly, the most common defect appeared to be the loss of STAT1.

    Article Title: Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush
    Article Snippet: Our data showed marked increases in STAT1 and STAT3 phosphorylation at 1, 3, and 6 hours after axonal injury, whereas the phosphorylation of STAT 2, 5, and 6 was not changed ( A), suggesting both STAT1 and STAT3 were activated after injury.

    Activity Assay:

    Article Title: Genome and Infection Characteristics of Human Parechovirus Type 1: The Interplay between Viral Infection and Type I Interferon Antiviral System
    Article Snippet: .. Previous study revealed enterovirus 71 2A protease reduces IFN receptor 1 activity to disrupt the activation of STAT1, STAT2, Jak1 and Tyk2 [ ]. .. These data may suggest future investigation of HPeV1 regulating type I IFN activity because we also found the HPeV1 can attenuate type I IFN downstream signaling and antiviral protein expression.

    Expressing:

    Article Title: Critical Role of the CXCL10/C-X-C Chemokine Receptor 3 Axis in Promoting Leukocyte Recruitment and Neuronal Injury during Traumatic Optic Neuropathy Induced by Optic Nerve Crush
    Article Snippet: .. At 4 hours after TON, Stattic blocks both STAT1 and STAT3 phosphorylation ( B), and significantly blocked TON-induced CXCL10 expression ( C). .. These results indicate that activation of STAT1/3 is involved in up-regulation of CXCL10 after axonal injury.

    SDS Page:

    Article Title: Rit Signaling Contributes to Interferon-γ-Induced Dendritic Retraction via p38 MAP Kinase Activation
    Article Snippet: .. Equal amounts of protein from each lysate were separated by SDS-PAGE (7%), transferred to PVDF membranes and reacted with antibodies that recognize both phosphorylated and non-phosphorylated STAT1 (Cell Signaling Technology, Beverly, MA) or with mAb that specifically recognizes STAT1 phosphorylated at Ser 727 (pSTAT1, Cell Signaling Technology). .. Blots were also probed using antibodies specific for α–tubulin (Sigma).

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    Cell Signaling Technology Inc rabbit polyclonal antibodies against tyr 701 phosphorylated stat1
    <t>U-STAT1,</t> U-STAT2, and IRF9 form U-ISGF3, which binds to ISREs on the target gene promoters. hTERT-HME1 cells expressing high levels of U-STAT1, U-STAT2, and IRF9 without IFN stimulation were analysed by co-immunoprecipitation (Co-IP) and chromatin-immunoprecipitation (ChIP) assays. ( A ) Nuclear proteins were used for Co-IP with normal rabbit IgG or rabbit <t>polyclonal</t> antibodies against STAT1, STAT2, or IRF9. To stabilize protein–protein interactions, the nuclear fraction was treated with the cleavable cross-linker, dimethyl-3,3′-dithiobis-propinimidate (DTBP, lanes 1, 2, and 4). Mouse monoclonal antibodies against STAT1, STAT2, or IRF9 were used for the western method. The purity of the fractions was assessed by determining the levels of GAPDH (a cytoplasmic protein) and HDAC1 (a nuclear protein) in the input lysates by the western method. ( B , C ) Total protein lysates were cross-linked with 1% formaldehyde and the cell lysates were cross-linked with DTBP. Chromatin was sheared into
    Rabbit Polyclonal Antibodies Against Tyr 701 Phosphorylated Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antibodies against tyr 701 phosphorylated stat1/product/Cell Signaling Technology Inc
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    Stimulation with type I interferon (IFN) induces IFN-induced transmembrane (IFITM) protein expression in human macrophages. Monocyte-derived macrophages were incubated with the indicated IFNs for 24 hours or left untreated and expression of <t>STAT1,</t> phosphorylated STAT1, β-actin, IFITM1, and IFITM2/3 was assessed by Western blot analysis. As controls, 293T cells were transfected with empty plasmid or expression plasmids for IFITM1–3. Similar results were obtained in 3 separate experiments. Abbreviation: MDM, monocyte-derived macrophages.
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    Esophageal cells with mutant p53 R175H and POSTN reveal activation of the <t>STAT1</t> signaling pathway. ( a ) Venn diagram displaying the number of genes with significant differential expression between the compared groups. Gene expression data were generated with RNA isolated from dissected epithelia of EPC-hTERT-p53 R175H -POSTN cells grown in organotypic culture ( n =3) compared with EPC-hTERT-p53 R175H -neo cells ( n =3) as well as parental non-invading EPC-hTERT cells ( n =3). The blue circle (gene lists hTERT and p53 R175H ) represents genes differentially expressed between EPC-hTERT and EPC-hTERT-p53 R175H -neo (3121). The red circle (gene lists p53 R175H and POSTN) represents genes differentially expressed between EPC-hTERT-p53 R175H -neo and EPC-hTERT-p53 R175H -POSTN (1808). ( P
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    Treatment with the <t>STAT1</t> inhibitor fludarabine reduces renal inflammation and ameliorates glomerular lesions in Habu GN. (A) Western blots showing P-STAT1, STAT1, and MHC class II expression levels in glomeruli of Habu GN, which were treated with the STAT1 inhibitor fludarabine, on days 3 and 7; representative images and a summary of the normalized quantification are shown. (B) PAS-stained kidney tissues were examined in Habu GN treated with fludarabine on days 3 and 7. (C) Mesangiolysis index and glomerulosclerosis index in the fludarabine-treated Habu GN group compared with those in the control group on days 3 and 7. (D) CD4+ T cell infiltration was suppressed in the fludarabine-treated group compared with that in the control group on days 3 and 7. (E) Relative IFN- γ, TNF- α, IL-12A, IL-12B, IL-6, IL-17A. and IL-23A mRNA levels in the fludarabine-treated Habu GN glomeruli as determined by RT-PCR. Data are presented as the mean ± SEM. (F) Effects of fludarabine on plasma creatinine and blood urea nitrogen (BUN) levels measured in Habu GN on D3 and D7. Data are presented as the mean ± SEM ( n = 10 per group). The results are representative of three independent experiments. * P
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    Image Search Results


    U-STAT1, U-STAT2, and IRF9 form U-ISGF3, which binds to ISREs on the target gene promoters. hTERT-HME1 cells expressing high levels of U-STAT1, U-STAT2, and IRF9 without IFN stimulation were analysed by co-immunoprecipitation (Co-IP) and chromatin-immunoprecipitation (ChIP) assays. ( A ) Nuclear proteins were used for Co-IP with normal rabbit IgG or rabbit polyclonal antibodies against STAT1, STAT2, or IRF9. To stabilize protein–protein interactions, the nuclear fraction was treated with the cleavable cross-linker, dimethyl-3,3′-dithiobis-propinimidate (DTBP, lanes 1, 2, and 4). Mouse monoclonal antibodies against STAT1, STAT2, or IRF9 were used for the western method. The purity of the fractions was assessed by determining the levels of GAPDH (a cytoplasmic protein) and HDAC1 (a nuclear protein) in the input lysates by the western method. ( B , C ) Total protein lysates were cross-linked with 1% formaldehyde and the cell lysates were cross-linked with DTBP. Chromatin was sheared into

    Journal: The EMBO Journal

    Article Title: IFN?-dependent increases in STAT1, STAT2, and IRF9 mediate resistance to viruses and DNA damage

    doi: 10.1038/emboj.2013.203

    Figure Lengend Snippet: U-STAT1, U-STAT2, and IRF9 form U-ISGF3, which binds to ISREs on the target gene promoters. hTERT-HME1 cells expressing high levels of U-STAT1, U-STAT2, and IRF9 without IFN stimulation were analysed by co-immunoprecipitation (Co-IP) and chromatin-immunoprecipitation (ChIP) assays. ( A ) Nuclear proteins were used for Co-IP with normal rabbit IgG or rabbit polyclonal antibodies against STAT1, STAT2, or IRF9. To stabilize protein–protein interactions, the nuclear fraction was treated with the cleavable cross-linker, dimethyl-3,3′-dithiobis-propinimidate (DTBP, lanes 1, 2, and 4). Mouse monoclonal antibodies against STAT1, STAT2, or IRF9 were used for the western method. The purity of the fractions was assessed by determining the levels of GAPDH (a cytoplasmic protein) and HDAC1 (a nuclear protein) in the input lysates by the western method. ( B , C ) Total protein lysates were cross-linked with 1% formaldehyde and the cell lysates were cross-linked with DTBP. Chromatin was sheared into

    Article Snippet: Mouse monoclonal antibodies against STAT1 (BD Transduction), STAT2, IRF9 (ISGF3γ), and IRF1 (Santa Cruz Biotechnology), and rabbit polyclonal antibodies against Tyr 701-phosphorylated STAT1, Tyr 690-phophorylated STAT2 (Cell Signaling), and STAT1 (Upstate) were used for western analyses.

    Techniques: Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Chromatin Immunoprecipitation, Western Blot

    Stimulation with type I interferon (IFN) induces IFN-induced transmembrane (IFITM) protein expression in human macrophages. Monocyte-derived macrophages were incubated with the indicated IFNs for 24 hours or left untreated and expression of STAT1, phosphorylated STAT1, β-actin, IFITM1, and IFITM2/3 was assessed by Western blot analysis. As controls, 293T cells were transfected with empty plasmid or expression plasmids for IFITM1–3. Similar results were obtained in 3 separate experiments. Abbreviation: MDM, monocyte-derived macrophages.

    Journal: The Journal of Infectious Diseases

    Article Title: Interferon-Induced Transmembrane Protein–Mediated Inhibition of Host Cell Entry of Ebolaviruses

    doi: 10.1093/infdis/jiv255

    Figure Lengend Snippet: Stimulation with type I interferon (IFN) induces IFN-induced transmembrane (IFITM) protein expression in human macrophages. Monocyte-derived macrophages were incubated with the indicated IFNs for 24 hours or left untreated and expression of STAT1, phosphorylated STAT1, β-actin, IFITM1, and IFITM2/3 was assessed by Western blot analysis. As controls, 293T cells were transfected with empty plasmid or expression plasmids for IFITM1–3. Similar results were obtained in 3 separate experiments. Abbreviation: MDM, monocyte-derived macrophages.

    Article Snippet: To detect STAT1 expression and phosphorylation, a polyclonal rabbit anti-STAT1α/β antibody and a polyclonal rabbit anti-phospho-STAT1 (Tyr701) antibody (both 1:1000; Cell Signaling) were used, respectively.

    Techniques: Expressing, Derivative Assay, Incubation, Western Blot, Transfection, Plasmid Preparation

    Esophageal cells with mutant p53 R175H and POSTN reveal activation of the STAT1 signaling pathway. ( a ) Venn diagram displaying the number of genes with significant differential expression between the compared groups. Gene expression data were generated with RNA isolated from dissected epithelia of EPC-hTERT-p53 R175H -POSTN cells grown in organotypic culture ( n =3) compared with EPC-hTERT-p53 R175H -neo cells ( n =3) as well as parental non-invading EPC-hTERT cells ( n =3). The blue circle (gene lists hTERT and p53 R175H ) represents genes differentially expressed between EPC-hTERT and EPC-hTERT-p53 R175H -neo (3121). The red circle (gene lists p53 R175H and POSTN) represents genes differentially expressed between EPC-hTERT-p53 R175H -neo and EPC-hTERT-p53 R175H -POSTN (1808). ( P

    Journal: Oncogenesis

    Article Title: Periostin cooperates with mutant p53 to mediate invasion through the induction of STAT1 signaling in the esophageal tumor microenvironment

    doi: 10.1038/oncsis.2013.17

    Figure Lengend Snippet: Esophageal cells with mutant p53 R175H and POSTN reveal activation of the STAT1 signaling pathway. ( a ) Venn diagram displaying the number of genes with significant differential expression between the compared groups. Gene expression data were generated with RNA isolated from dissected epithelia of EPC-hTERT-p53 R175H -POSTN cells grown in organotypic culture ( n =3) compared with EPC-hTERT-p53 R175H -neo cells ( n =3) as well as parental non-invading EPC-hTERT cells ( n =3). The blue circle (gene lists hTERT and p53 R175H ) represents genes differentially expressed between EPC-hTERT and EPC-hTERT-p53 R175H -neo (3121). The red circle (gene lists p53 R175H and POSTN) represents genes differentially expressed between EPC-hTERT-p53 R175H -neo and EPC-hTERT-p53 R175H -POSTN (1808). ( P

    Article Snippet: For immunohistochemistry, rabbit polyclonal POSTN (Abcam, ab 14041) and rabbit monoclonal phospho-STAT1 (Tyr701; Cell Signaling) were used.

    Techniques: Mutagenesis, Activation Assay, Expressing, Generated, Isolation

    STAT1 knockdown in EPC-hTERT-p53 R175H -POSTN and transformed EPC-hTERT-EGFR-p53 R175H cells show decrease in invasion. ( a ) Western blot confirming knockdown total STAT1 and STAT1 phosphorylation in invasive EPC-hTERT-p53 R175H -POSTN and in transformed, genetically engineered EPC-hTERT-EGFR-p53 R175H cells using two independent shRNAs directed against STAT1 and non-specific shRNAs as controls (A and B represent independently generated cell lines with the same genotype). GAPDH was used as a loading control. ( b ) Transwell Boyden Chamber invasion assay of EPC-hTERT-p53 R175H -POSTN-shSTAT1-A and -B and EPC-hTERT-EGFR-p53 R175H -shSTAT1-A and -B cells compared with control EPC-hTERT-p53 R175H -POSTN-shNS-A and -B and EPC-hTERT-EGFR-p53 R175H -shNS-A and -B cells. Bar graphs represent fold changes±s.e.m. * P

    Journal: Oncogenesis

    Article Title: Periostin cooperates with mutant p53 to mediate invasion through the induction of STAT1 signaling in the esophageal tumor microenvironment

    doi: 10.1038/oncsis.2013.17

    Figure Lengend Snippet: STAT1 knockdown in EPC-hTERT-p53 R175H -POSTN and transformed EPC-hTERT-EGFR-p53 R175H cells show decrease in invasion. ( a ) Western blot confirming knockdown total STAT1 and STAT1 phosphorylation in invasive EPC-hTERT-p53 R175H -POSTN and in transformed, genetically engineered EPC-hTERT-EGFR-p53 R175H cells using two independent shRNAs directed against STAT1 and non-specific shRNAs as controls (A and B represent independently generated cell lines with the same genotype). GAPDH was used as a loading control. ( b ) Transwell Boyden Chamber invasion assay of EPC-hTERT-p53 R175H -POSTN-shSTAT1-A and -B and EPC-hTERT-EGFR-p53 R175H -shSTAT1-A and -B cells compared with control EPC-hTERT-p53 R175H -POSTN-shNS-A and -B and EPC-hTERT-EGFR-p53 R175H -shNS-A and -B cells. Bar graphs represent fold changes±s.e.m. * P

    Article Snippet: For immunohistochemistry, rabbit polyclonal POSTN (Abcam, ab 14041) and rabbit monoclonal phospho-STAT1 (Tyr701; Cell Signaling) were used.

    Techniques: Transformation Assay, Western Blot, Generated, Invasion Assay

    Inducible knockdown of POSTN in ESCC xenograft tumors display decreased p53 expression and STAT1 activation. ( a ) Phospho-STAT1(Tyr701) expression by immunohistochemistry of tumors formed in vivo by subcutaneous injection of HCE4 cancer cells stably transfected with either lentiviral doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) vectors. Left panels represent tumors that were not induced with doxycycline (DOX), and right panels represent tumors induced with doxycycline. Bar=100 μℳ. ( b ) Phospho-STAT1(Tyr701) expression by immunohistochemistry of tumors formed in vivo by subcutaneous injection of TE-11 cancer cells stably transfected with either lentiviral doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) vectors. Left panels represent tumors that were not induced with doxycycline, and right panels represent tumors induced with doxycycline. Bar=100 μℳ. ( c ) Western blot analysis of STAT1 and p53 expression in four pairs of lysates isolated from HCE4 xenograft tumors transduced with doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) with or without doxycycline treatment. Immunoblotting for POSTN expression to confirm doxycycline induced knockdown. GAPDH was used as a loading control. ( d ) Western blot analysis of STAT1 and p53 expression in four pairs of lysates isolated from TE-11 xenograft tumors transduced with doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) with or without doxycycline treatment. Immunoblotting for POSTN expression to confirm doxycycline induced knockdown. GAPDH was used as a loading control.

    Journal: Oncogenesis

    Article Title: Periostin cooperates with mutant p53 to mediate invasion through the induction of STAT1 signaling in the esophageal tumor microenvironment

    doi: 10.1038/oncsis.2013.17

    Figure Lengend Snippet: Inducible knockdown of POSTN in ESCC xenograft tumors display decreased p53 expression and STAT1 activation. ( a ) Phospho-STAT1(Tyr701) expression by immunohistochemistry of tumors formed in vivo by subcutaneous injection of HCE4 cancer cells stably transfected with either lentiviral doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) vectors. Left panels represent tumors that were not induced with doxycycline (DOX), and right panels represent tumors induced with doxycycline. Bar=100 μℳ. ( b ) Phospho-STAT1(Tyr701) expression by immunohistochemistry of tumors formed in vivo by subcutaneous injection of TE-11 cancer cells stably transfected with either lentiviral doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) vectors. Left panels represent tumors that were not induced with doxycycline, and right panels represent tumors induced with doxycycline. Bar=100 μℳ. ( c ) Western blot analysis of STAT1 and p53 expression in four pairs of lysates isolated from HCE4 xenograft tumors transduced with doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) with or without doxycycline treatment. Immunoblotting for POSTN expression to confirm doxycycline induced knockdown. GAPDH was used as a loading control. ( d ) Western blot analysis of STAT1 and p53 expression in four pairs of lysates isolated from TE-11 xenograft tumors transduced with doxycycline-inducible non-specific targeting shRNA (shNS) or shRNA specific to periostin (shPOSTN) with or without doxycycline treatment. Immunoblotting for POSTN expression to confirm doxycycline induced knockdown. GAPDH was used as a loading control.

    Article Snippet: For immunohistochemistry, rabbit polyclonal POSTN (Abcam, ab 14041) and rabbit monoclonal phospho-STAT1 (Tyr701; Cell Signaling) were used.

    Techniques: Expressing, Activation Assay, Immunohistochemistry, In Vivo, Injection, Stable Transfection, Transfection, shRNA, Western Blot, Isolation, Transduction

    Treatment with the STAT1 inhibitor fludarabine reduces renal inflammation and ameliorates glomerular lesions in Habu GN. (A) Western blots showing P-STAT1, STAT1, and MHC class II expression levels in glomeruli of Habu GN, which were treated with the STAT1 inhibitor fludarabine, on days 3 and 7; representative images and a summary of the normalized quantification are shown. (B) PAS-stained kidney tissues were examined in Habu GN treated with fludarabine on days 3 and 7. (C) Mesangiolysis index and glomerulosclerosis index in the fludarabine-treated Habu GN group compared with those in the control group on days 3 and 7. (D) CD4+ T cell infiltration was suppressed in the fludarabine-treated group compared with that in the control group on days 3 and 7. (E) Relative IFN- γ, TNF- α, IL-12A, IL-12B, IL-6, IL-17A. and IL-23A mRNA levels in the fludarabine-treated Habu GN glomeruli as determined by RT-PCR. Data are presented as the mean ± SEM. (F) Effects of fludarabine on plasma creatinine and blood urea nitrogen (BUN) levels measured in Habu GN on D3 and D7. Data are presented as the mean ± SEM ( n = 10 per group). The results are representative of three independent experiments. * P

    Journal: Frontiers in Immunology

    Article Title: Suppressor of Cytokine Signaling-1/STAT1 Regulates Renal Inflammation in Mesangial Proliferative Glomerulonephritis Models

    doi: 10.3389/fimmu.2018.01982

    Figure Lengend Snippet: Treatment with the STAT1 inhibitor fludarabine reduces renal inflammation and ameliorates glomerular lesions in Habu GN. (A) Western blots showing P-STAT1, STAT1, and MHC class II expression levels in glomeruli of Habu GN, which were treated with the STAT1 inhibitor fludarabine, on days 3 and 7; representative images and a summary of the normalized quantification are shown. (B) PAS-stained kidney tissues were examined in Habu GN treated with fludarabine on days 3 and 7. (C) Mesangiolysis index and glomerulosclerosis index in the fludarabine-treated Habu GN group compared with those in the control group on days 3 and 7. (D) CD4+ T cell infiltration was suppressed in the fludarabine-treated group compared with that in the control group on days 3 and 7. (E) Relative IFN- γ, TNF- α, IL-12A, IL-12B, IL-6, IL-17A. and IL-23A mRNA levels in the fludarabine-treated Habu GN glomeruli as determined by RT-PCR. Data are presented as the mean ± SEM. (F) Effects of fludarabine on plasma creatinine and blood urea nitrogen (BUN) levels measured in Habu GN on D3 and D7. Data are presented as the mean ± SEM ( n = 10 per group). The results are representative of three independent experiments. * P

    Article Snippet: Sections were used for immunohistochemical analysis with the following antibodies: proliferating cell nuclear antigen (PCNA; Cell Signaling Technology), anti-CD4 (Abcam), P-STAT1 (Cell Signaling Technology), and rabbit IgG polyclonal-Isotype Control (Abcam).

    Techniques: Western Blot, Expressing, Staining, Reverse Transcription Polymerase Chain Reaction

    SOCS-1 inhibits IFN-γ-induced CIITA promoter IV activity and gene expression via STAT1 regulation. (A) Quantitative RT-PCR analysis of CIITA mRNA expression in MMCs transfected with SOCS1 plasmids or treated with fludarabine for 48 h incubated in the absence or presence of IFN-γ for 48 h. (B) MMCs were cotransfected with the CIITA promoter and SOCS1 plasmid or treated with fludarabine for 48 h in the absence or presence of IFN-γ for 48 h prior to analyze for luciferase activity as indicated in the Materials and Methods. Statistical significance was determined relative to the negative control. Data are presented as the mean ± SEM ( n = 3) of at least three experiments. * P

    Journal: Frontiers in Immunology

    Article Title: Suppressor of Cytokine Signaling-1/STAT1 Regulates Renal Inflammation in Mesangial Proliferative Glomerulonephritis Models

    doi: 10.3389/fimmu.2018.01982

    Figure Lengend Snippet: SOCS-1 inhibits IFN-γ-induced CIITA promoter IV activity and gene expression via STAT1 regulation. (A) Quantitative RT-PCR analysis of CIITA mRNA expression in MMCs transfected with SOCS1 plasmids or treated with fludarabine for 48 h incubated in the absence or presence of IFN-γ for 48 h. (B) MMCs were cotransfected with the CIITA promoter and SOCS1 plasmid or treated with fludarabine for 48 h in the absence or presence of IFN-γ for 48 h prior to analyze for luciferase activity as indicated in the Materials and Methods. Statistical significance was determined relative to the negative control. Data are presented as the mean ± SEM ( n = 3) of at least three experiments. * P

    Article Snippet: Sections were used for immunohistochemical analysis with the following antibodies: proliferating cell nuclear antigen (PCNA; Cell Signaling Technology), anti-CD4 (Abcam), P-STAT1 (Cell Signaling Technology), and rabbit IgG polyclonal-Isotype Control (Abcam).

    Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Transfection, Incubation, Plasmid Preparation, Luciferase, Negative Control

    Fludarabine inhibits IFN-γ-induced MHC class II expression and chemokine production. (A) Western blot analysis of P-STAT1 and STAT1 in IFN-γ-treated MMCs in the absence or presence of fludarabine for 48 h. (B) Quantitative RT-PCR analysis of MHC class II mRNA expression in IFN-γ-stimulated MMCs in the absence or presence of fludarabine for 48 h. (C) Representative Western blots showing the levels of MHC class II in IFN-γ-stimulated mesangial cells in the absence or presence of fludarabine for 48 h. (D) Relative levels of IP-10 and Mig mRNA in IFN-γ-treated MMCs in the absence or presence of fludarabine for 48 h as determined by RT-PCR. Data are presented as the mean ± SEM ( n = 3). The results are representative of three independent experiments. * P

    Journal: Frontiers in Immunology

    Article Title: Suppressor of Cytokine Signaling-1/STAT1 Regulates Renal Inflammation in Mesangial Proliferative Glomerulonephritis Models

    doi: 10.3389/fimmu.2018.01982

    Figure Lengend Snippet: Fludarabine inhibits IFN-γ-induced MHC class II expression and chemokine production. (A) Western blot analysis of P-STAT1 and STAT1 in IFN-γ-treated MMCs in the absence or presence of fludarabine for 48 h. (B) Quantitative RT-PCR analysis of MHC class II mRNA expression in IFN-γ-stimulated MMCs in the absence or presence of fludarabine for 48 h. (C) Representative Western blots showing the levels of MHC class II in IFN-γ-stimulated mesangial cells in the absence or presence of fludarabine for 48 h. (D) Relative levels of IP-10 and Mig mRNA in IFN-γ-treated MMCs in the absence or presence of fludarabine for 48 h as determined by RT-PCR. Data are presented as the mean ± SEM ( n = 3). The results are representative of three independent experiments. * P

    Article Snippet: Sections were used for immunohistochemical analysis with the following antibodies: proliferating cell nuclear antigen (PCNA; Cell Signaling Technology), anti-CD4 (Abcam), P-STAT1 (Cell Signaling Technology), and rabbit IgG polyclonal-Isotype Control (Abcam).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

    SOCS1 and STAT1 expression in MsGN models. (A) Immunofluorescence analysis was performed to determine SOCS1 expression in Thy1.1 and Habu GN glomeruli (Thy1.1: magnification × 200; Habu: magnification × 400). (B) SOCS1 protein expression in Thy1.1 and Habu GN glomeruli. Quantitative analysis of SOCS1 protein expression in glomeruli of MsGN models. (C) Quantitative RT-PCR analysis of SOCS1 mRNA expression in glomeruli of MsGN models. (D) Immunohistochemistry analysis of P-STAT1 in Thy1.1 and Habu GN kidney sections. Representative micrographs and quantification of positive cells in glomeruli are shown (original magnification, × 400). (E) Western blot analysis of P-STAT1 and STAT1 expression in Thy1.1 and Habu GN glomeruli; representative images and the summary of the normalized quantification are shown. Data are presented as the mean ± SEM ( n = 10 per group). The results are representative of three independent experiments. * P

    Journal: Frontiers in Immunology

    Article Title: Suppressor of Cytokine Signaling-1/STAT1 Regulates Renal Inflammation in Mesangial Proliferative Glomerulonephritis Models

    doi: 10.3389/fimmu.2018.01982

    Figure Lengend Snippet: SOCS1 and STAT1 expression in MsGN models. (A) Immunofluorescence analysis was performed to determine SOCS1 expression in Thy1.1 and Habu GN glomeruli (Thy1.1: magnification × 200; Habu: magnification × 400). (B) SOCS1 protein expression in Thy1.1 and Habu GN glomeruli. Quantitative analysis of SOCS1 protein expression in glomeruli of MsGN models. (C) Quantitative RT-PCR analysis of SOCS1 mRNA expression in glomeruli of MsGN models. (D) Immunohistochemistry analysis of P-STAT1 in Thy1.1 and Habu GN kidney sections. Representative micrographs and quantification of positive cells in glomeruli are shown (original magnification, × 400). (E) Western blot analysis of P-STAT1 and STAT1 expression in Thy1.1 and Habu GN glomeruli; representative images and the summary of the normalized quantification are shown. Data are presented as the mean ± SEM ( n = 10 per group). The results are representative of three independent experiments. * P

    Article Snippet: Sections were used for immunohistochemical analysis with the following antibodies: proliferating cell nuclear antigen (PCNA; Cell Signaling Technology), anti-CD4 (Abcam), P-STAT1 (Cell Signaling Technology), and rabbit IgG polyclonal-Isotype Control (Abcam).

    Techniques: Expressing, Immunofluorescence, Quantitative RT-PCR, Immunohistochemistry, Western Blot

    SOCS1 expression inhibits IFN-γ-induced MHC class II expression and chemokine production. (A) Flow cytometric analysis of MHC class II expression in IFN-γ-stimulated mesangial cells. (B) Quantitative RT-PCR analysis of MHC class II mRNA expression in IFN-γ-stimulated MMCs transfected with SOCS1 plasmids. (C) MMCs transfected with SOCS1 plasmids were incubated in the absence or presence of IFN-γ for 48 h. Representative Western blots show MHC class II levels in IFN-γ-stimulated mesangial cells. (D) Relative IP-10 and Mig mRNA levels in IFN-γ-treated MMCs transfected with SOCS1 plasmids as determined by RT-PCR. (E) Western blot analysis of P-STAT1 and STAT1 expression in SOCS1 plasmid-transfected MMCs incubated in the absence or presence of IFN-γ for 48 h; representative images and the summary of the normalized quantification are shown. Data are presented as the mean ± SEM ( n = 3). The results are representative of three independent experiments. * P

    Journal: Frontiers in Immunology

    Article Title: Suppressor of Cytokine Signaling-1/STAT1 Regulates Renal Inflammation in Mesangial Proliferative Glomerulonephritis Models

    doi: 10.3389/fimmu.2018.01982

    Figure Lengend Snippet: SOCS1 expression inhibits IFN-γ-induced MHC class II expression and chemokine production. (A) Flow cytometric analysis of MHC class II expression in IFN-γ-stimulated mesangial cells. (B) Quantitative RT-PCR analysis of MHC class II mRNA expression in IFN-γ-stimulated MMCs transfected with SOCS1 plasmids. (C) MMCs transfected with SOCS1 plasmids were incubated in the absence or presence of IFN-γ for 48 h. Representative Western blots show MHC class II levels in IFN-γ-stimulated mesangial cells. (D) Relative IP-10 and Mig mRNA levels in IFN-γ-treated MMCs transfected with SOCS1 plasmids as determined by RT-PCR. (E) Western blot analysis of P-STAT1 and STAT1 expression in SOCS1 plasmid-transfected MMCs incubated in the absence or presence of IFN-γ for 48 h; representative images and the summary of the normalized quantification are shown. Data are presented as the mean ± SEM ( n = 3). The results are representative of three independent experiments. * P

    Article Snippet: Sections were used for immunohistochemical analysis with the following antibodies: proliferating cell nuclear antigen (PCNA; Cell Signaling Technology), anti-CD4 (Abcam), P-STAT1 (Cell Signaling Technology), and rabbit IgG polyclonal-Isotype Control (Abcam).

    Techniques: Expressing, Flow Cytometry, Quantitative RT-PCR, Transfection, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation