murine smad7 Search Results


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  • 99
    New England Biolabs m mulv reverse transcriptase
    M Mulv Reverse Transcriptase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3547 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC murine c2c12 myoblasts
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Murine C2c12 Myoblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Stratagene quickhyb solution
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Quickhyb Solution, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 461 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher ptripz shp21
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Ptripz Shp21, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore puromycin
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Puromycin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 41497 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher moloney murine leukemia virus
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Moloney Murine Leukemia Virus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1856 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cancer Therapeutics CRC colorectal cancer
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Colorectal Cancer, supplied by Cancer Therapeutics CRC, used in various techniques. Bioz Stars score: 92/100, based on 820 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millennium Pharmaceuticals xenopus smad8 cdna
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Xenopus Smad8 Cdna, supplied by Millennium Pharmaceuticals, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Cell Signaling Technology Inc oct 4a
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Oct 4a, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    PeproTech recombinant il6
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Recombinant Il6, supplied by PeproTech, used in various techniques. Bioz Stars score: 96/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore insulin
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
    Insulin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 21861 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen rneasy micro kit
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Millipore tgf α
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    R&D Systems vegf165
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Thermo Fisher alexa flour conjugated antibodies
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Genechem pgcl vector
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Thermo Fisher abi prism 7500
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Thermo Fisher mouse monoclonal anti ham56 igm
    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in <t>C2C12</t> cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.
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    Santa Cruz Biotechnology smad2
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    Abcam smurf2
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    Santa Cruz Biotechnology goat polyclonal anti smad7 igg
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    Biogenex antigen retrieval
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    Santa Cruz Biotechnology smad7
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    Millipore kynurenic acid
    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) <t>SMAD2</t> and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.
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    YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in C2C12 cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.

    Journal: Molecular and Cellular Biology

    Article Title: Nuclear Factor YY1 Inhibits Transforming Growth Factor ?- and Bone Morphogenetic Protein-Induced Cell Differentiation

    doi: 10.1128/MCB.23.13.4494-4510.2003

    Figure Lengend Snippet: YY1 represses TGF-β and BMP immediate-early gene responses. (A) YY1 represses PAI-1 induction by TGF-β1. Northern analysis of PAI-1 mRNA from Mv1Lu cells mock treated (−) or treated (+) for 24 h with 10 ng of TGF-β1 per ml in the absence of YY1 (−) or in the presence of increasing doses of YY1 (Ad-YY1) (triangle; MOI of 20 or 100) or control GFP (Ad-GFP) (MOI of 20 or 100) adenovirus is shown. Sample loading and blotting efficiency was monitored with β-actin mRNA on the same blot. (B) YY1 represses Id-1 induction by TGF-β1. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells infected with the indicated MOI of recombinant adenoviruses and then treated with vehicle (−) or 2.5 ng of TGF-β1 per ml (+) for 1 h is shown. Arrows mark the positions of endogenous Id-1, adenoviral YY1, adenoviral Flag-Smad4 (F-Smad4), endogenous phosphorylated Smad2 (Smad2-P), and endogenous β-catenin, which serves as a loading and blotting efficiency control. (C) YY1 represses Id-1 induction by BMP-7. Immunoblot analysis of endogenous Id-1 expression in MDA-MB-468 cells is as for panel B but in response to 300 ng of BMP-7 per ml. Infection conditions were as for panel B, and arrows mark the same proteins as in panel B except for endogenous phosphorylated Smad1 (Smad1-P) instead of Smad2. Asterisks mark nonspecific protein bands. (D) YY1 represses PAI-1 promoter induction by TGF-β1. PAI-1 (800)-luciferase assays were performed in HepG2 cells with increasing amounts of YY1 plasmid (triangle; 0.1 to 1.0 μg) in the absence (−) or presence (+) of overnight stimulation with 2.5 ng of TGF-β1 per ml. The averaged data for normalized reporter activity are shown with standard errors derived from triplicate transfections repeated at least twice independently. (E) YY1 represses the Smad-specific 12xCAGA promoter. The assays were similar to those for panel D, with the 12xCAGA-luciferase reporter in HepG2 cells after overnight stimulation with 2.5 ng of TGF-β1 per ml. Data are reported as in panel D. (F) YY1 represses the Smad-specific Id1-BRE 2 promoter. The assays were similar to those for panel D, with the Id1-BRE 2 -luciferase reporter in C2C12 cells after overnight stimulation with 100 ng of BMP-6 per ml. Data are reported as in panel D.

    Article Snippet: Murine C2C12 myoblasts, murine mammary epithelial NMuMG cells, mink lung epithelial Mv1Lu cells, monkey kidney COS-7 cells, human embryonic kidney 293T cells, human mammary MDA-MB-468 cells, and human hepatoma HepG2 cells were obtained and cultured according to protocols from the American Type Culture Collection.

    Techniques: Northern Blot, Expressing, Multiple Displacement Amplification, Infection, Recombinant, Luciferase, Plasmid Preparation, Activity Assay, Derivative Assay, Transfection

    YY1 interacts physically with Smads in vitro. (A) Schematic representation of Smad proteins with their conserved MH1, the DNA-binding β-hairpin loop (black boxes), MH2, and the less conserved linker sequences. The strength of Smad protein interaction with YY1 is quantified based on the data in panels B and C. (B) Endogenous YY1 from C2C12 cells interacts with the indicated GST-Smad fusions with different relative affinities (lanes 2 to 5). A total C2C12 lysate immunoblot is shown for comparison (lane 6). Ponceau S staining of the immunoblot shows the immobilized GST-Smads and GST protein. (C) YY1 interacts with the MH1 domain of Smad4. Experiment similar to those for panel B were performed with endogenous YY1 from HaCaT cells. Immobilized GST fusion proteins are shown in a Coomassie brilliant blue (CBB) stain of the gel. In panels B and C, arrows mark the specific protein bands.

    Journal: Molecular and Cellular Biology

    Article Title: Nuclear Factor YY1 Inhibits Transforming Growth Factor ?- and Bone Morphogenetic Protein-Induced Cell Differentiation

    doi: 10.1128/MCB.23.13.4494-4510.2003

    Figure Lengend Snippet: YY1 interacts physically with Smads in vitro. (A) Schematic representation of Smad proteins with their conserved MH1, the DNA-binding β-hairpin loop (black boxes), MH2, and the less conserved linker sequences. The strength of Smad protein interaction with YY1 is quantified based on the data in panels B and C. (B) Endogenous YY1 from C2C12 cells interacts with the indicated GST-Smad fusions with different relative affinities (lanes 2 to 5). A total C2C12 lysate immunoblot is shown for comparison (lane 6). Ponceau S staining of the immunoblot shows the immobilized GST-Smads and GST protein. (C) YY1 interacts with the MH1 domain of Smad4. Experiment similar to those for panel B were performed with endogenous YY1 from HaCaT cells. Immobilized GST fusion proteins are shown in a Coomassie brilliant blue (CBB) stain of the gel. In panels B and C, arrows mark the specific protein bands.

    Article Snippet: Murine C2C12 myoblasts, murine mammary epithelial NMuMG cells, mink lung epithelial Mv1Lu cells, monkey kidney COS-7 cells, human embryonic kidney 293T cells, human mammary MDA-MB-468 cells, and human hepatoma HepG2 cells were obtained and cultured according to protocols from the American Type Culture Collection.

    Techniques: In Vitro, Binding Assay, Staining

    Ectopic expression of YY1 inhibits TGF-β- and BMP-induced transdifferentiation. (A) Top panels, YY1 inhibits mammary NMuMG cell EMT induced by TGF-β1. NMuMG cells were infected with Ad-GFP (MOI, 25) or Ad-YY1 (MOI, 10) adenovirus, treated for 24 h with vehicle (control) or with 2.5 ng of TGF-β1 per ml, and subjected to actin direct fluorescence microscopy. Bottom panels, YY1 inhibits NMuMG EMT induced by the TGF-β type I receptor. EMT induced by caALK-5 receptor (MOI, 60) is blocked by coinfection with the YY1 virus at an MOI of 20 (Ad-YY1) or with the Smad7 virus at an MOI of 20 (Ad-Smad7). Bar, 20 μm. (B) YY1 blocks BMP-7-induced preosteoblast differentiation of C2C12 myoblasts. In situ ALP staining of C2C12 cells treated with vehicle (control) or 300 ng of BMP-7 per ml for 3 days after infection with Ad-GFP (MOI, 20) or Ad-YY1 (MOI, 4) virus is shown. Bar, 80 μm.

    Journal: Molecular and Cellular Biology

    Article Title: Nuclear Factor YY1 Inhibits Transforming Growth Factor ?- and Bone Morphogenetic Protein-Induced Cell Differentiation

    doi: 10.1128/MCB.23.13.4494-4510.2003

    Figure Lengend Snippet: Ectopic expression of YY1 inhibits TGF-β- and BMP-induced transdifferentiation. (A) Top panels, YY1 inhibits mammary NMuMG cell EMT induced by TGF-β1. NMuMG cells were infected with Ad-GFP (MOI, 25) or Ad-YY1 (MOI, 10) adenovirus, treated for 24 h with vehicle (control) or with 2.5 ng of TGF-β1 per ml, and subjected to actin direct fluorescence microscopy. Bottom panels, YY1 inhibits NMuMG EMT induced by the TGF-β type I receptor. EMT induced by caALK-5 receptor (MOI, 60) is blocked by coinfection with the YY1 virus at an MOI of 20 (Ad-YY1) or with the Smad7 virus at an MOI of 20 (Ad-Smad7). Bar, 20 μm. (B) YY1 blocks BMP-7-induced preosteoblast differentiation of C2C12 myoblasts. In situ ALP staining of C2C12 cells treated with vehicle (control) or 300 ng of BMP-7 per ml for 3 days after infection with Ad-GFP (MOI, 20) or Ad-YY1 (MOI, 4) virus is shown. Bar, 80 μm.

    Article Snippet: Murine C2C12 myoblasts, murine mammary epithelial NMuMG cells, mink lung epithelial Mv1Lu cells, monkey kidney COS-7 cells, human embryonic kidney 293T cells, human mammary MDA-MB-468 cells, and human hepatoma HepG2 cells were obtained and cultured according to protocols from the American Type Culture Collection.

    Techniques: Expressing, Infection, Fluorescence, Microscopy, In Situ, ALP Assay, Staining

    Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) SMAD2 and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.

    Journal: Frontiers in Pharmacology

    Article Title: Proteomics-Based Characterization of miR-574-5p Decoy to CUGBP1 Suggests Specificity for mPGES-1 Regulation in Human Lung Cancer Cells

    doi: 10.3389/fphar.2020.00196

    Figure Lengend Snippet: Identification of novel CUGBP1 targets. Western blots of IL-1β-stimulated A549 cells with ΔCUGBP1 showed increased levels of (A) NDUFS2, (B) mPGES-1, (C) SMAD2 and (D) SMAD3 compared to control. β-Actin was used as loading control. Fold induction is given as mean (+SEM) of three independent experiments, t -test *** p ≤ 0.001. (E,F) RIP with antibodies against CUGBP1 or IgG (mock) showed enrichment of bound mRNAs quantified via q RT-PCR. (E) cJUN was used as positive control, COX-2 as negative control. (F) mRNAs of NDUFS2, mPGES-1, SMAD2 and CUGBP1 were enriched in CUGBP1-IP, while SMAD3 mRNA showed no enrichment. Relative enrichment normalized to IgG is given as mean (+SEM) from three independent experiments. t -test * p ≤ 0.05; ** p ≤ 0.005. RIP: RNA immunoprecipitation.

    Article Snippet: SMAD2, SMAD3 and SMAD4 were detected in the proteomics study and expression changes were successfully validated using Western blot analysis.

    Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control, Immunoprecipitation