c-jun Santa Cruz Biotechnology Search Results


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  • 95
    Millipore c jun
    C Jun, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc c jun
    C Jun, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 3023 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Santa Cruz Biotechnology anti jun b santa cruz
    Effect of EGF on the interaction between <t>Sp1</t> oligonucleotides and c-Jun/Sp1. Nuclear extracts from EGF-treated cells were prepared and subjected to the assay for binding of the c-Jun/Sp1 complex to Sp1 consensus sites. The 32 P-radiolabeled Sp1 oligonucleotide (●) and Sp1 mutant SPM (□) were used as a probe for binding, respectively. For background control, protein A-agarose was used to substitute <t>anti-c-Jun</t> antibody–agarose conjugate in the assay (■). Values are means ± SEM of three determinations.
    Anti Jun B Santa Cruz, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 77/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Santa Cruz Biotechnology gst c jun
    Effect of Tax on <t>JNK1</t> enzymatic activity. (A) 293 cells were transfected with 500 ng of κB-TATA-luciferase, 500 ng of FLAG-JNK1, and 50 ng of 6RZ and either 0.125, 0.5, or 2 μg of an N-terminally truncated but constitutively active form of MEKK1 or 1 μg of either Myc-TRAF2 or HTLV-1 Tax. Finally, 293 cells transfected with reporters only were treated for 30 min and 6 h with 100 ng of TNF-α per ml. Cell lysates were prepared to determine luciferase activity and β-galactosidase activity and were used for immunoprecipitation with anti-FLAG M2 antibody. Anti-FLAG M2 immunoprecipitates from each transfection were subjected to in vitro kinase assays in the presence of <t>GST–c-Jun(1-79)</t> as an exogenous substrate. In vitro kinase reactions were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were subsequently probed with anti-FLAG M2 antibody to establish levels of immunoprecipitated FLAG-JNK1. (B) κB-luciferase activity was measured in the same cell lysates as used for panel A and normalized to β-galactosidase activity to control for differences in transfection efficiency. The data are presented as relative luciferase activity based on arbitrary light units.
    Gst C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 81/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology c jun ap1
    Effect of Tax on <t>JNK1</t> enzymatic activity. (A) 293 cells were transfected with 500 ng of κB-TATA-luciferase, 500 ng of FLAG-JNK1, and 50 ng of 6RZ and either 0.125, 0.5, or 2 μg of an N-terminally truncated but constitutively active form of MEKK1 or 1 μg of either Myc-TRAF2 or HTLV-1 Tax. Finally, 293 cells transfected with reporters only were treated for 30 min and 6 h with 100 ng of TNF-α per ml. Cell lysates were prepared to determine luciferase activity and β-galactosidase activity and were used for immunoprecipitation with anti-FLAG M2 antibody. Anti-FLAG M2 immunoprecipitates from each transfection were subjected to in vitro kinase assays in the presence of <t>GST–c-Jun(1-79)</t> as an exogenous substrate. In vitro kinase reactions were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were subsequently probed with anti-FLAG M2 antibody to establish levels of immunoprecipitated FLAG-JNK1. (B) κB-luciferase activity was measured in the same cell lysates as used for panel A and normalized to β-galactosidase activity to control for differences in transfection efficiency. The data are presented as relative luciferase activity based on arbitrary light units.
    C Jun Ap1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 75/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology nonphospho c jun
    Effect of Tax on <t>JNK1</t> enzymatic activity. (A) 293 cells were transfected with 500 ng of κB-TATA-luciferase, 500 ng of FLAG-JNK1, and 50 ng of 6RZ and either 0.125, 0.5, or 2 μg of an N-terminally truncated but constitutively active form of MEKK1 or 1 μg of either Myc-TRAF2 or HTLV-1 Tax. Finally, 293 cells transfected with reporters only were treated for 30 min and 6 h with 100 ng of TNF-α per ml. Cell lysates were prepared to determine luciferase activity and β-galactosidase activity and were used for immunoprecipitation with anti-FLAG M2 antibody. Anti-FLAG M2 immunoprecipitates from each transfection were subjected to in vitro kinase assays in the presence of <t>GST–c-Jun(1-79)</t> as an exogenous substrate. In vitro kinase reactions were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were subsequently probed with anti-FLAG M2 antibody to establish levels of immunoprecipitated FLAG-JNK1. (B) κB-luciferase activity was measured in the same cell lysates as used for panel A and normalized to β-galactosidase activity to control for differences in transfection efficiency. The data are presented as relative luciferase activity based on arbitrary light units.
    Nonphospho C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology phosphorylated c jun
    Involvement of activation of transcription factors, NF-κB and /AP-1, in IL-1β-induced IL-36γ expression. (A) IL-1β induced IκBα phosphorylation and degradation in colonic myofibroblasts. The cells were stimulated with IL-1β (10 ng/ml), and phosphorylated IκBα were detected by Western blotting. (B) IL-1β induced activation of NF-κB and c-Jun AP-1. The cells were stimulated with IL-1β (10 ng/ml), and NF-κB p65 and phosphorylated c-Jun were detected by immunocytochemistory. Reacted antibodies against NF-κB p65 were visualized by FITC (green fluorescence)-labeled second antibody. Reacted antibodies against phosphorylated c-Jun were detected by a DyLight ®  594 (red fluorescence)-labeled secondary antibodies. Nucleus was stained by DAPI (blue). (C) The cells were stimulated with IL-1β (10 ng/ml) or medium alone for 15 min and nuclear proteins were extracted. NF-κB p65 and phosphorylated c-Jun in nuclear extracts were detected by immunoblot. (D) Effects of silencing of NF-κB p65 and c-Jun AP-1on IL-1β-induced IL-36γ expression. The cells were transfected with control siRNA, the siRNA specific for NF-κB p65 and/or c-Jun AP-1, and incubated for 24h. IL-36γ mRNA expression was analyzed by real-time PCR. IL-36γ mRNA expression was expressed relative to the β-actin mRNA expression (mean ± SD from 4 different experiments). *P
    Phosphorylated C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti c jun
    Differential association of AP-1 components and <t>ERβ</t> with the PTPRO promoter in the presence of E2 and tamoxifen. A, Whole-cell extracts from Hs578t, 48R, and MCF-7 cells were subjected to Western blot analysis with <t>anti-c-Fos</t> and anti-c-Jun antibody.
    Anti C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 1024 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Santa Cruz Biotechnology phosphor c jun
    Endothelial CYP2J2 overexpression increases PI3K/ AKT activation and altered ERK1/2 and c-Jun/JNK activation after BCCAO after BCCAO. (A) Immunoblotting (upper) of p-AKT ,total AKT and PI3K and densitometric analysis (bottom) using nuclear extract isolated from cortex 24 hours after BCCAO. p-AKT and PI3K protein upregulation after ischemia was augmented in Tie2-CYP2J2 Tr (2J2 Tr) mice, C26 inhibited the level of p-AKT and PI3K up regulated in 2J2 Tr mice. (B, C, D) Phospho-ERK1/2, ERK1/2, <t>phosphor-c-Jun,</t> c-Jun, phosphor-JNK and total-JNK expression in brain homogenates was assessed by immunblotting and densitometric analysis. Phospho-ERK1/2 (panels B) was significantly increased after BCCAO in Tie-CYP2J2-Tr (2J2 Tr) mice compared to WT. Phosphorylation of c-Jun (panels C) was increased after ischemia in WT but not in Tie2-CYP2J2 Tr brains. Phosphorylation of JNK (panels D) was increased after ischemia in WT but not in Tie2-CYP2J2-Tr brains. However, applied C26 reduced these effect. Blots are representative of three independent experiments. * p
    Phosphor C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 82/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology c jun sc 45
    Endothelial CYP2J2 overexpression increases PI3K/ AKT activation and altered ERK1/2 and c-Jun/JNK activation after BCCAO after BCCAO. (A) Immunoblotting (upper) of p-AKT ,total AKT and PI3K and densitometric analysis (bottom) using nuclear extract isolated from cortex 24 hours after BCCAO. p-AKT and PI3K protein upregulation after ischemia was augmented in Tie2-CYP2J2 Tr (2J2 Tr) mice, C26 inhibited the level of p-AKT and PI3K up regulated in 2J2 Tr mice. (B, C, D) Phospho-ERK1/2, ERK1/2, <t>phosphor-c-Jun,</t> c-Jun, phosphor-JNK and total-JNK expression in brain homogenates was assessed by immunblotting and densitometric analysis. Phospho-ERK1/2 (panels B) was significantly increased after BCCAO in Tie-CYP2J2-Tr (2J2 Tr) mice compared to WT. Phosphorylation of c-Jun (panels C) was increased after ischemia in WT but not in Tie2-CYP2J2 Tr brains. Phosphorylation of JNK (panels D) was increased after ischemia in WT but not in Tie2-CYP2J2-Tr brains. However, applied C26 reduced these effect. Blots are representative of three independent experiments. * p
    C Jun Sc 45, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Santa Cruz Biotechnology sirna c jun
    TGF-β3-induced autophagy contributed to increased MUC5AC production by activating the AP1. (a-b) 16HBE cells were transduced with <t>ATG5-siRNA</t> lentivirus (a) and BECN1-siRNA lentivirus (b), respectively. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (c-d) 16HBE cells were treated with TGF-β3 in the presence of 3-MA (c) or Baf A1 (d). Then, the expression of phospho-c-Jun and c-Jun were detected using western blot assay. (e) 16HBE cells were transfected with <t>Smad2/3-siRNA.</t> After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (f) 16HBE cells were transfected with c-Jun-siRNA, after treating with TGF-β3 (10 ng/ml) for 24 h, and then phospho-c-Jun and c-Jun were detected by western blot. (g) Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells were transfected with c-Jun-siRNA. (h) Quantitation of fluorescence intensity of MUC5AC (each group n = 10 images for quantification). (i) 16HBE cells were transfected with c-Jun-siRNA. Real-time PCR was performed to detect the expression of MUC5AC gene after treated with TGF-β3 (10 ng/ml). Data are representative of the three independent experiments and are presented as means ± s.d . *P
    Sirna C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 76/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology human c jun
    Recombinant full-length dimeric c-Jun-containing human AP-1 complexes are all active in binding the consensus TRE sequence found in the human cyclin D1 gene. A , shown is a schematic drawing of human c-Jun and Fos family proteins tagged at the N terminus
    Human C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology c jun mabs
    Recombinant full-length dimeric c-Jun-containing human AP-1 complexes are all active in binding the consensus TRE sequence found in the human cyclin D1 gene. A , shown is a schematic drawing of human c-Jun and Fos family proteins tagged at the N terminus
    C Jun Mabs, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Santa Cruz Biotechnology anti c jun polyclonal
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    Anti C Jun Polyclonal, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 79/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology c jun specific antibody
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    C Jun Specific Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 83/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti c jun ap1
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    Anti C Jun Ap1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Santa Cruz Biotechnology km 1 against c jun
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    Km 1 Against C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Santa Cruz Biotechnology mouse antiphospho c jun
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    Mouse Antiphospho C Jun, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Santa Cruz Biotechnology c jun ser 63 73
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
    C Jun Ser 63 73, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 87/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Santa Cruz Biotechnology rabbit α c jun
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
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    Santa Cruz Biotechnology c jun sc44x
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
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    Santa Cruz Biotechnology c jun sc 1694
    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with <t>polyclonal</t> c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal <t>anti-c-Jun</t> antibodies (upper panel) or anti-Pin1 antibodies (lower panel).
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    Santa Cruz Biotechnology rabbit anti c jun
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Santa Cruz Biotechnology monoclonal c jun antibody
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Santa Cruz Biotechnology phospho c jun ser63
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Santa Cruz Biotechnology anti c jun sc 822
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Santa Cruz Biotechnology serine 63 phosphorylated c jun
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Santa Cruz Biotechnology anti c jun h 79
    C-Jun was required for TNC-induced <t>MMP9</t> expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and <t>anti-c-Jun</t> (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P
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    Image Search Results


    Effect of EGF on the interaction between Sp1 oligonucleotides and c-Jun/Sp1. Nuclear extracts from EGF-treated cells were prepared and subjected to the assay for binding of the c-Jun/Sp1 complex to Sp1 consensus sites. The 32 P-radiolabeled Sp1 oligonucleotide (●) and Sp1 mutant SPM (□) were used as a probe for binding, respectively. For background control, protein A-agarose was used to substitute anti-c-Jun antibody–agarose conjugate in the assay (■). Values are means ± SEM of three determinations.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Functional interaction between c-Jun and promoter factor Sp1 in epidermal growth factor-induced gene expression of human 12(S)-lipoxygenase

    doi:

    Figure Lengend Snippet: Effect of EGF on the interaction between Sp1 oligonucleotides and c-Jun/Sp1. Nuclear extracts from EGF-treated cells were prepared and subjected to the assay for binding of the c-Jun/Sp1 complex to Sp1 consensus sites. The 32 P-radiolabeled Sp1 oligonucleotide (●) and Sp1 mutant SPM (□) were used as a probe for binding, respectively. For background control, protein A-agarose was used to substitute anti-c-Jun antibody–agarose conjugate in the assay (■). Values are means ± SEM of three determinations.

    Article Snippet: Polyclonal antibodies against c-Jun and Sp1, protein A-agarose and agarose conjugated to Sp1 or c-Jun antibodies were from Santa Cruz Biotechnology.

    Techniques: Binding Assay, Mutagenesis

    Binding of c-Jun and Sp1 in cells overexpressing c-Jun or Ha-ras. Cells were transfected with a different amount of pRSVjun or pSV2ras by the lipofection method. After the change of Opti-MEM medium to 3 ml of fresh culture medium in a 6-cm plastic dish, cells were incubated for an additional 36 h. Expression of c-Jun protein ( A ) and the coimmunoprecipitated c-Jun/Sp1 complex by using anti-Sp1 antibodies ( B and C ) was analyzed by Western blot with anti-c-Jun and anti-Sp1 antibodies.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Functional interaction between c-Jun and promoter factor Sp1 in epidermal growth factor-induced gene expression of human 12(S)-lipoxygenase

    doi:

    Figure Lengend Snippet: Binding of c-Jun and Sp1 in cells overexpressing c-Jun or Ha-ras. Cells were transfected with a different amount of pRSVjun or pSV2ras by the lipofection method. After the change of Opti-MEM medium to 3 ml of fresh culture medium in a 6-cm plastic dish, cells were incubated for an additional 36 h. Expression of c-Jun protein ( A ) and the coimmunoprecipitated c-Jun/Sp1 complex by using anti-Sp1 antibodies ( B and C ) was analyzed by Western blot with anti-c-Jun and anti-Sp1 antibodies.

    Article Snippet: Polyclonal antibodies against c-Jun and Sp1, protein A-agarose and agarose conjugated to Sp1 or c-Jun antibodies were from Santa Cruz Biotechnology.

    Techniques: Binding Assay, Transfection, Incubation, Expressing, Western Blot

    Effect of c-Jun dominant negative mutant on c-Jun/Sp1 interaction in pRSVjun-treated cells. Cells were transfected with 2 μg of pRSVjun and a different amount of c-Jun dominant negative vector TAM-67 by the lipofection method. After the change of Opti-MEM medium to 3 ml of fresh culture medium in a 6-cm plastic dish, cells were incubated for an additional 36 h. Expression of c-Jun and TAM-67 proteins ( A ) and the coimmunoprecipitated c-Jun/Sp1 and TAM-67/Sp1 complex by using anti-Sp1 antibodies ( B ) was analyzed by Western blot with anti-c-Jun and anti-Sp1 antibodies. pcDNA3.1 was used as a vector to adjust for the same amount of plasmids in each experiment.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Functional interaction between c-Jun and promoter factor Sp1 in epidermal growth factor-induced gene expression of human 12(S)-lipoxygenase

    doi:

    Figure Lengend Snippet: Effect of c-Jun dominant negative mutant on c-Jun/Sp1 interaction in pRSVjun-treated cells. Cells were transfected with 2 μg of pRSVjun and a different amount of c-Jun dominant negative vector TAM-67 by the lipofection method. After the change of Opti-MEM medium to 3 ml of fresh culture medium in a 6-cm plastic dish, cells were incubated for an additional 36 h. Expression of c-Jun and TAM-67 proteins ( A ) and the coimmunoprecipitated c-Jun/Sp1 and TAM-67/Sp1 complex by using anti-Sp1 antibodies ( B ) was analyzed by Western blot with anti-c-Jun and anti-Sp1 antibodies. pcDNA3.1 was used as a vector to adjust for the same amount of plasmids in each experiment.

    Article Snippet: Polyclonal antibodies against c-Jun and Sp1, protein A-agarose and agarose conjugated to Sp1 or c-Jun antibodies were from Santa Cruz Biotechnology.

    Techniques: Dominant Negative Mutation, Transfection, Plasmid Preparation, Incubation, Expressing, Western Blot

    (A) Coimmunoprecipitation assays were performed by immunoprecipitation (IP) with anti-c-Jun and blotting with anti-PU.1 and anti-c-Jun. To control for nuclear protein extract concentrations, levels of the nuclear protein TATA binding protein (TBP) were examined. (B) For ChIP assays, isolated lysates were immunoprecipitated with either anti-PU.1 or anti-c-Jun, followed by amplification of purified DNA fragments using primers specific for the c- Jun promoter or Mcsfr promoter or by antiacetylated H4, followed by amplification of purified DNA fragments using primers specific for the HPRT promoter.

    Journal: Molecular and Cellular Biology

    Article Title: Increased c-Jun Expression and Reduced GATA2 Expression Promote Aberrant Monocytic Differentiation Induced by Activating PTPN11 Mutants ▿ Mutants ▿ §

    doi: 10.1128/MCB.01330-08

    Figure Lengend Snippet: (A) Coimmunoprecipitation assays were performed by immunoprecipitation (IP) with anti-c-Jun and blotting with anti-PU.1 and anti-c-Jun. To control for nuclear protein extract concentrations, levels of the nuclear protein TATA binding protein (TBP) were examined. (B) For ChIP assays, isolated lysates were immunoprecipitated with either anti-PU.1 or anti-c-Jun, followed by amplification of purified DNA fragments using primers specific for the c- Jun promoter or Mcsfr promoter or by antiacetylated H4, followed by amplification of purified DNA fragments using primers specific for the HPRT promoter.

    Article Snippet: For coimmunoprecipitation assays, 1 mg of nuclear protein extract was incubated with anti-c-Jun antibody (H-79; Santa Cruz Biotechnology) and protein A Sepharose beads (CL-4B; GE Healthcare) for 2 h at 4°C.

    Techniques: Immunoprecipitation, Binding Assay, Chromatin Immunoprecipitation, Isolation, Amplification, Purification

    Effect of HNE on the phosphorylation of c-Myc. HBE1 cells were treated with 10 μM HNE for the indicated time points. Whole cell extracts were collected and subjected to immunobloting with anti p-c-Myc. c-Myc was used as loading control and identified with anti-c-Myc antibody. The figure shows a representative experiment and graph reported as mean ± S.E. out of three independent experiments.

    Journal: IUBMB life

    Article Title: c-Myc is a Nrf2-interacting protein that negatively regulates phase II genes through their electrophile responsive elements

    doi: 10.1002/iub.314

    Figure Lengend Snippet: Effect of HNE on the phosphorylation of c-Myc. HBE1 cells were treated with 10 μM HNE for the indicated time points. Whole cell extracts were collected and subjected to immunobloting with anti p-c-Myc. c-Myc was used as loading control and identified with anti-c-Myc antibody. The figure shows a representative experiment and graph reported as mean ± S.E. out of three independent experiments.

    Article Snippet: Anti-c-Myc, anti-p-c-Myc, anti-p-c-Jun, anti-c-Jun, anti- Nrf2 and anti-lamin antibodies were from Santa Cruz Biotechnology, as well as c-Myc- and Nrf2- siRNA (Santa Cruz, CA).

    Techniques: Western Blot

    Interaction of c-Myc with either Nrf2 or p-c-Jun at both basal and induced conditions. HBE1 cells were treated with 10 μM HNE or vehicle control for 3 hours. Nuclear proteins were immunoprecipitated with the indicated antibodies: A. c-Myc. B. Nrf2. C. p-c-Jun, and visualized by Western blot analysis with anti Nrf2, anti c-Myc, anti p-c-Jun, anti p-c-Myc or anti c-Jun antibodies. Lamin was used as loading control and identified with anti-lamin antibody. Expression levels of the indicated proteins in the whole cell extract are shown in total lysate (lower panels).

    Journal: IUBMB life

    Article Title: c-Myc is a Nrf2-interacting protein that negatively regulates phase II genes through their electrophile responsive elements

    doi: 10.1002/iub.314

    Figure Lengend Snippet: Interaction of c-Myc with either Nrf2 or p-c-Jun at both basal and induced conditions. HBE1 cells were treated with 10 μM HNE or vehicle control for 3 hours. Nuclear proteins were immunoprecipitated with the indicated antibodies: A. c-Myc. B. Nrf2. C. p-c-Jun, and visualized by Western blot analysis with anti Nrf2, anti c-Myc, anti p-c-Jun, anti p-c-Myc or anti c-Jun antibodies. Lamin was used as loading control and identified with anti-lamin antibody. Expression levels of the indicated proteins in the whole cell extract are shown in total lysate (lower panels).

    Article Snippet: Anti-c-Myc, anti-p-c-Myc, anti-p-c-Jun, anti-c-Jun, anti- Nrf2 and anti-lamin antibodies were from Santa Cruz Biotechnology, as well as c-Myc- and Nrf2- siRNA (Santa Cruz, CA).

    Techniques: Immunoprecipitation, Western Blot, Expressing

    IL-1 and OSM activate c-fos expression and AP-1 binding to the AP-1 element from the PAI-1 gene A. Human astrocytes were stimulated with IL-1 or OSM for 1 hour and then nuclear extracts were prepared and binding analyzed using a PAI-1-AP-1 double stranded oligonucleotide. Extracts were incubated with anti-c-fos, anti-c-jun antibodies or normal rabbit serum (NRS) for 10 minutes when indicated. Asterisk indicates position of the IL-1/OSM activated band. Arrows marks super-shifted complexes. B. Cells were stimulated with IL-1 or OSM for indicated time periods and nuclear extracts were prepared. Extracts were incubated with anti-c-fos antibodies and binding analyzed using a PAI-1-AP-1 double stranded oligonucleotide. C. Astrocytes were stimulated with IL-1 or OSM for indicated time periods and lysates were prepared under denaturing conditions. Lysates were separated by SDS-PAGE and transferred to nitrocellulose membrane. C-fos was detected by anti-c-fos antibodies and complexes were detected by ECL.

    Journal: Journal of neurochemistry

    Article Title: Mechanism of Plasminogen Activator Inhibitor-1 regulation by Oncostatin M and Interleukin-1 in human astrocytes

    doi:

    Figure Lengend Snippet: IL-1 and OSM activate c-fos expression and AP-1 binding to the AP-1 element from the PAI-1 gene A. Human astrocytes were stimulated with IL-1 or OSM for 1 hour and then nuclear extracts were prepared and binding analyzed using a PAI-1-AP-1 double stranded oligonucleotide. Extracts were incubated with anti-c-fos, anti-c-jun antibodies or normal rabbit serum (NRS) for 10 minutes when indicated. Asterisk indicates position of the IL-1/OSM activated band. Arrows marks super-shifted complexes. B. Cells were stimulated with IL-1 or OSM for indicated time periods and nuclear extracts were prepared. Extracts were incubated with anti-c-fos antibodies and binding analyzed using a PAI-1-AP-1 double stranded oligonucleotide. C. Astrocytes were stimulated with IL-1 or OSM for indicated time periods and lysates were prepared under denaturing conditions. Lysates were separated by SDS-PAGE and transferred to nitrocellulose membrane. C-fos was detected by anti-c-fos antibodies and complexes were detected by ECL.

    Article Snippet: The polyclonal anti-c-fos and anti-c-jun antisera were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).

    Techniques: Expressing, Binding Assay, Incubation, SDS Page

    Cot kinase enhances c-Jun expression. (A) NIH 3T3 cells were transfected with a wild-type Cot (a and b)-, CotKR (c and d)-, or MEKK1 (e and f)-expressing vector (0.5 μg). At 24 h after transfection, cells were transferred into serum-free medium for an additional 24 h, fixed, and then analyzed by double immunofluorescence with specific anti-c-Jun (b, d, and f) and anti-Cot/Tpl2 (a and c) or anti-MEKK1 (e) antibodies. (B) NIH 3T3 cells were transfected with the reporter plasmid pJLuc, which encodes a luc gene driven by the wild-type c- jun promoter. These cells were also cotransfected with expression vectors (1 μg) encoding the MEKK1 Cot, and Raf proteins, as indicated. At 24 h after transfection, cells were transferred to serum-free medium for an additional 24 h, lysed, and then analyzed for luciferase activity as described in Materials and Methods. Luciferase activity was normalized with respect to that of vector-transfected cells, whose value was taken as 1. β-Galactosidase activity resulting from the cotransfection of a pCDNAIII β-galactosidase expression vector was used to normalize the luciferase values to the corresponding transfection efficiencies. The results shown are averages ± the standard errors of triplicate samples from a typical experiment. Similar results were obtained in three independent experiments. c, control.

    Journal: Molecular and Cellular Biology

    Article Title: Multiple Mitogen-Activated Protein Kinase Signaling Pathways Connect the Cot Oncoprotein to the c-jun Promoter and to Cellular Transformation

    doi:

    Figure Lengend Snippet: Cot kinase enhances c-Jun expression. (A) NIH 3T3 cells were transfected with a wild-type Cot (a and b)-, CotKR (c and d)-, or MEKK1 (e and f)-expressing vector (0.5 μg). At 24 h after transfection, cells were transferred into serum-free medium for an additional 24 h, fixed, and then analyzed by double immunofluorescence with specific anti-c-Jun (b, d, and f) and anti-Cot/Tpl2 (a and c) or anti-MEKK1 (e) antibodies. (B) NIH 3T3 cells were transfected with the reporter plasmid pJLuc, which encodes a luc gene driven by the wild-type c- jun promoter. These cells were also cotransfected with expression vectors (1 μg) encoding the MEKK1 Cot, and Raf proteins, as indicated. At 24 h after transfection, cells were transferred to serum-free medium for an additional 24 h, lysed, and then analyzed for luciferase activity as described in Materials and Methods. Luciferase activity was normalized with respect to that of vector-transfected cells, whose value was taken as 1. β-Galactosidase activity resulting from the cotransfection of a pCDNAIII β-galactosidase expression vector was used to normalize the luciferase values to the corresponding transfection efficiencies. The results shown are averages ± the standard errors of triplicate samples from a typical experiment. Similar results were obtained in three independent experiments. c, control.

    Article Snippet: The cells were incubated with an anti-c-Jun MAb (Santa Cruz Biotecnology, Inc.) and an anti-Tpl2/Cot (Santa Cruz Biotechnology, Inc.) or anti-MEKK1 antibody for 1 h and washed three times with PBS and then with a 1:100 dilution of fluorescein-conjugated goat anti-mouse F(ab′)2 IgG and tetramethyl rhodamine isocyanate-conjugated anti-rabbit F(ab′)2 IgG antibodies (Jackson ImmunoResearch Laboratories, Inc.).

    Techniques: Expressing, Transfection, Plasmid Preparation, Immunofluorescence, Luciferase, Activity Assay, Cotransfection

    Figure 5 AP‐1 mediates cyclooxygenase (COX)‐2 induction by CD and DC. (A) SEG‐1 cells were treated with 200 μM of CD and DC for 16–24 h, and immunoblots were performed with c‐Jun/AP‐1,

    Journal: Gut

    Article Title: COX-2 induction by unconjugated bile acids involves reactive oxygen species-mediated signalling pathways in Barrett's oesophagus and oesophageal adenocarcinoma

    doi: 10.1136/gut.2007.121244

    Figure Lengend Snippet: Figure 5 AP‐1 mediates cyclooxygenase (COX)‐2 induction by CD and DC. (A) SEG‐1 cells were treated with 200 μM of CD and DC for 16–24 h, and immunoblots were performed with c‐Jun/AP‐1,

    Article Snippet: Dulbecco's modified Eagle medium (DMEM) and fetal bovine serum (FBS) (Life Technologies, Inc, New York, USA); LY294002 and U0126 (Calbiochem, San Diego, CA, USA); conjugated and unconjugated bile acids, 2,7‐dichlorodihydrofluorescein diacetate (DCHF‐DA); N‐acetyl‐L‐cysteine (NAC) and sodium formate (Sigma, St Louis, USA); ECL‐chemiluminescence reagents for western blots (Amersham‐Pharmacia, New Jersey, USA); total and phosphorylated ERK1/2 antibodies, total and phosphorylated AKT antibodies, and phosphorylated CREB antibody (Cell Signaling Technology, Beverly, USA); COX‐2 monoclonal antibody (Cayman Chemical, Ann Arbor, USA) and c‐Jun/AP‐1 antibody (Santa Cruz Biotechnology, Inc, Santa Cruz, USA).

    Techniques: Western Blot

    Effect of Tax on JNK1 enzymatic activity. (A) 293 cells were transfected with 500 ng of κB-TATA-luciferase, 500 ng of FLAG-JNK1, and 50 ng of 6RZ and either 0.125, 0.5, or 2 μg of an N-terminally truncated but constitutively active form of MEKK1 or 1 μg of either Myc-TRAF2 or HTLV-1 Tax. Finally, 293 cells transfected with reporters only were treated for 30 min and 6 h with 100 ng of TNF-α per ml. Cell lysates were prepared to determine luciferase activity and β-galactosidase activity and were used for immunoprecipitation with anti-FLAG M2 antibody. Anti-FLAG M2 immunoprecipitates from each transfection were subjected to in vitro kinase assays in the presence of GST–c-Jun(1-79) as an exogenous substrate. In vitro kinase reactions were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were subsequently probed with anti-FLAG M2 antibody to establish levels of immunoprecipitated FLAG-JNK1. (B) κB-luciferase activity was measured in the same cell lysates as used for panel A and normalized to β-galactosidase activity to control for differences in transfection efficiency. The data are presented as relative luciferase activity based on arbitrary light units.

    Journal: Molecular and Cellular Biology

    Article Title: Human T-Cell Leukemia Virus Type 1 Tax Induction of NF-?B Involves Activation of the I?B Kinase ? (IKK?) and IKK? Cellular Kinases

    doi:

    Figure Lengend Snippet: Effect of Tax on JNK1 enzymatic activity. (A) 293 cells were transfected with 500 ng of κB-TATA-luciferase, 500 ng of FLAG-JNK1, and 50 ng of 6RZ and either 0.125, 0.5, or 2 μg of an N-terminally truncated but constitutively active form of MEKK1 or 1 μg of either Myc-TRAF2 or HTLV-1 Tax. Finally, 293 cells transfected with reporters only were treated for 30 min and 6 h with 100 ng of TNF-α per ml. Cell lysates were prepared to determine luciferase activity and β-galactosidase activity and were used for immunoprecipitation with anti-FLAG M2 antibody. Anti-FLAG M2 immunoprecipitates from each transfection were subjected to in vitro kinase assays in the presence of GST–c-Jun(1-79) as an exogenous substrate. In vitro kinase reactions were subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were subsequently probed with anti-FLAG M2 antibody to establish levels of immunoprecipitated FLAG-JNK1. (B) κB-luciferase activity was measured in the same cell lysates as used for panel A and normalized to β-galactosidase activity to control for differences in transfection efficiency. The data are presented as relative luciferase activity based on arbitrary light units.

    Article Snippet: Cotransfection of an N-terminally truncated MEKK1 mutant ( ) which is constitutively active led to substantial activation of JNK1 as assessed by phosphorylation of GST–c-Jun(1-79) substrate in vitro (Fig. A).

    Techniques: Activity Assay, Transfection, Luciferase, Immunoprecipitation, In Vitro, SDS Page

    Involvement of activation of transcription factors, NF-κB and /AP-1, in IL-1β-induced IL-36γ expression. (A) IL-1β induced IκBα phosphorylation and degradation in colonic myofibroblasts. The cells were stimulated with IL-1β (10 ng/ml), and phosphorylated IκBα were detected by Western blotting. (B) IL-1β induced activation of NF-κB and c-Jun AP-1. The cells were stimulated with IL-1β (10 ng/ml), and NF-κB p65 and phosphorylated c-Jun were detected by immunocytochemistory. Reacted antibodies against NF-κB p65 were visualized by FITC (green fluorescence)-labeled second antibody. Reacted antibodies against phosphorylated c-Jun were detected by a DyLight ®  594 (red fluorescence)-labeled secondary antibodies. Nucleus was stained by DAPI (blue). (C) The cells were stimulated with IL-1β (10 ng/ml) or medium alone for 15 min and nuclear proteins were extracted. NF-κB p65 and phosphorylated c-Jun in nuclear extracts were detected by immunoblot. (D) Effects of silencing of NF-κB p65 and c-Jun AP-1on IL-1β-induced IL-36γ expression. The cells were transfected with control siRNA, the siRNA specific for NF-κB p65 and/or c-Jun AP-1, and incubated for 24h. IL-36γ mRNA expression was analyzed by real-time PCR. IL-36γ mRNA expression was expressed relative to the β-actin mRNA expression (mean ± SD from 4 different experiments). *P

    Journal: PLoS ONE

    Article Title: Interleukin (IL)-1β Is a Strong Inducer of IL-36γ Expression in Human Colonic Myofibroblasts

    doi: 10.1371/journal.pone.0138423

    Figure Lengend Snippet: Involvement of activation of transcription factors, NF-κB and /AP-1, in IL-1β-induced IL-36γ expression. (A) IL-1β induced IκBα phosphorylation and degradation in colonic myofibroblasts. The cells were stimulated with IL-1β (10 ng/ml), and phosphorylated IκBα were detected by Western blotting. (B) IL-1β induced activation of NF-κB and c-Jun AP-1. The cells were stimulated with IL-1β (10 ng/ml), and NF-κB p65 and phosphorylated c-Jun were detected by immunocytochemistory. Reacted antibodies against NF-κB p65 were visualized by FITC (green fluorescence)-labeled second antibody. Reacted antibodies against phosphorylated c-Jun were detected by a DyLight ® 594 (red fluorescence)-labeled secondary antibodies. Nucleus was stained by DAPI (blue). (C) The cells were stimulated with IL-1β (10 ng/ml) or medium alone for 15 min and nuclear proteins were extracted. NF-κB p65 and phosphorylated c-Jun in nuclear extracts were detected by immunoblot. (D) Effects of silencing of NF-κB p65 and c-Jun AP-1on IL-1β-induced IL-36γ expression. The cells were transfected with control siRNA, the siRNA specific for NF-κB p65 and/or c-Jun AP-1, and incubated for 24h. IL-36γ mRNA expression was analyzed by real-time PCR. IL-36γ mRNA expression was expressed relative to the β-actin mRNA expression (mean ± SD from 4 different experiments). *P

    Article Snippet: Antibodies against phosphorylated c-Jun, NF-κB p65, phosphorylated IκBα were purchased from Santa Cruz.

    Techniques: Activation Assay, Expressing, Western Blot, Fluorescence, Labeling, Staining, Transfection, Incubation, Real-time Polymerase Chain Reaction

    Investigation of the phosphorylation status of MAPK downstream effectors in LMP2A-expressing clones. (A) Antibodies that specifically react with phospho-c-Jun were used to investigate the phosphorylation of c-Jun in stably expressing clones (stable) and transient transfectants (trans) in LMP2A-expressing (2A) and vector control (V) cells. UV-treated (200 J/m 2 ) 293 cells (UV) served as the positive controls for phospho-c-Jun induction. (B) Antibodies that specifically react with phospho-ATF2 were used to investigate the phosphorylation of ATF2 in LMP2A-expressing (2A) and vector control (V) cells. The detection of tubulin served as an internal control of protein amounts. The numbers to the left of each panel represent molecular mass in kilodaltons.

    Journal: Journal of Virology

    Article Title: Epstein-Barr Virus Latent Membrane Protein 2A Regulates c-Jun Protein through Extracellular Signal-Regulated Kinase

    doi: 10.1128/JVI.76.18.9556-9561.2002

    Figure Lengend Snippet: Investigation of the phosphorylation status of MAPK downstream effectors in LMP2A-expressing clones. (A) Antibodies that specifically react with phospho-c-Jun were used to investigate the phosphorylation of c-Jun in stably expressing clones (stable) and transient transfectants (trans) in LMP2A-expressing (2A) and vector control (V) cells. UV-treated (200 J/m 2 ) 293 cells (UV) served as the positive controls for phospho-c-Jun induction. (B) Antibodies that specifically react with phospho-ATF2 were used to investigate the phosphorylation of ATF2 in LMP2A-expressing (2A) and vector control (V) cells. The detection of tubulin served as an internal control of protein amounts. The numbers to the left of each panel represent molecular mass in kilodaltons.

    Article Snippet: The levels of phospho-c-Jun (Ser 63), and phospho-ATF2 (activating transcription factor 2) were detected with anti-phospho-c-Jun (Santa Cruz, Santa Cruz, Calif.) and anti-phospho-ATF2 antibodies (Cell Signaling Technology).

    Techniques: Expressing, Stable Transfection, Plasmid Preparation

    CEP-1347 treatment decreases JNK activity and c-Jun phosphorylation in MCF-7 and LCC9 cells Cells were treated with vehicle or 100 nM CEP-1347 for 48 h. Cellular lysates were prepared and analyzed by western blotting using the indicated antibodies. Actin was used as loading control. (A) Levels of phospho-c-Jun in vehicle and CEP-1347 treated cells. Upper : representative blots. Lower : quantitative analysis of p-c-Jun levels, normalized to β-actin levels. Results represent the mean +/− SD of three independent experiments. **p

    Journal: Oncotarget

    Article Title: Targeting mixed lineage kinases in ER-positive breast cancer cells leads to G2/M cell cycle arrest and apoptosis

    doi:

    Figure Lengend Snippet: CEP-1347 treatment decreases JNK activity and c-Jun phosphorylation in MCF-7 and LCC9 cells Cells were treated with vehicle or 100 nM CEP-1347 for 48 h. Cellular lysates were prepared and analyzed by western blotting using the indicated antibodies. Actin was used as loading control. (A) Levels of phospho-c-Jun in vehicle and CEP-1347 treated cells. Upper : representative blots. Lower : quantitative analysis of p-c-Jun levels, normalized to β-actin levels. Results represent the mean +/− SD of three independent experiments. **p

    Article Snippet: Antibodies against Bax (2772), PARP (9542), phospho-ERK (9106), phospho-JNK (9255), and phospho-p38 (9216), were from Cell Signaling Biotechnology, Inc. Antibodies to phospho-c-Jun (sc-822), c-Jun (sc-1694), ERK1/2 (sc-93), JNK1/2 (sc-475), p38 (sc-535) and NF-κB p65 (sc-372-G), were from Santa Cruz Biotechnology.

    Techniques: Activity Assay, Western Blot

    Stretch stimulation activates select members of the AP-1 family of transcription factors. A: Nuclear extracts (NE) prepared from control or stretch-stimulated pBSMCs were used in an EMSA and assessed for binding to a radiolabeled oligonucleotide carrying a consensus AP-1–binding sequence motif. AP-1–DNA complex formation was detected in both basal and stretch-stimulated pBSMCs (lanes 2 and 9), which was competed out by increasing amounts of unlabeled homologous competitor oligonucleotide [wild type (WT), lanes 3 to 5 and 10 to 12] but not by competitor oligonucleotides carrying a mutated AP-1 site (mutant, lanes 6 to 8 and 13 to 15). Lane 1 is the free radiolabeled probe (no nuclear extract added) FP, free probe. B: The AP-1 complex (lanes 2 and 5) was supershifted (SS) in the presence of antibody to phospho-c-Jun (lanes 4 and 7) but not by an isotype-matched IgG control (lanes 3 and 6). Lane 1, free probe. C: Nuclear extracts from pBSMCs subjected to cyclic stretch for 2 hours or not (control) were assayed using a transcription factor ELISA to determine the DNA-binding activity of AP-1 proteins. The graph shows AP-1 binding expressed as a percentage of control for each of seven AP-1 subunits, and data are representative of at least two independent experiments. Increased binding of c-Jun, Fos, and FosB proteins to a consensus AP-1 oligonucleotide was observed in stretch-treated cells relative to controls. D: Protein lysates from pBSMCs (control or stretched for 24 hours) were immunoblotted with the indicated antibodies. E: Lysates from stretched (24 hours) or control pBSMCs were immunoprecipitated with FosB or isotype-matched IgG control antibodies and immunoblotted for c-Jun. An increase in Jun-FosB interaction is observed on stretch. The decrease in mobility of the c-Jun band is likely because of protein phosphorylation, consistent with D . Nonspecific interaction with the heavy chain of IgG in the sepharose beads used for immunoprecipitation (IP) is observed (IgG H ). All data shown are representative of at least three independent experiments.

    Journal: The American Journal of Pathology

    Article Title: FosB Regulates Stretch-Induced Expression of Extracellular Matrix Proteins in Smooth Muscle

    doi: 10.1016/j.ajpath.2011.08.034

    Figure Lengend Snippet: Stretch stimulation activates select members of the AP-1 family of transcription factors. A: Nuclear extracts (NE) prepared from control or stretch-stimulated pBSMCs were used in an EMSA and assessed for binding to a radiolabeled oligonucleotide carrying a consensus AP-1–binding sequence motif. AP-1–DNA complex formation was detected in both basal and stretch-stimulated pBSMCs (lanes 2 and 9), which was competed out by increasing amounts of unlabeled homologous competitor oligonucleotide [wild type (WT), lanes 3 to 5 and 10 to 12] but not by competitor oligonucleotides carrying a mutated AP-1 site (mutant, lanes 6 to 8 and 13 to 15). Lane 1 is the free radiolabeled probe (no nuclear extract added) FP, free probe. B: The AP-1 complex (lanes 2 and 5) was supershifted (SS) in the presence of antibody to phospho-c-Jun (lanes 4 and 7) but not by an isotype-matched IgG control (lanes 3 and 6). Lane 1, free probe. C: Nuclear extracts from pBSMCs subjected to cyclic stretch for 2 hours or not (control) were assayed using a transcription factor ELISA to determine the DNA-binding activity of AP-1 proteins. The graph shows AP-1 binding expressed as a percentage of control for each of seven AP-1 subunits, and data are representative of at least two independent experiments. Increased binding of c-Jun, Fos, and FosB proteins to a consensus AP-1 oligonucleotide was observed in stretch-treated cells relative to controls. D: Protein lysates from pBSMCs (control or stretched for 24 hours) were immunoblotted with the indicated antibodies. E: Lysates from stretched (24 hours) or control pBSMCs were immunoprecipitated with FosB or isotype-matched IgG control antibodies and immunoblotted for c-Jun. An increase in Jun-FosB interaction is observed on stretch. The decrease in mobility of the c-Jun band is likely because of protein phosphorylation, consistent with D . Nonspecific interaction with the heavy chain of IgG in the sepharose beads used for immunoprecipitation (IP) is observed (IgG H ). All data shown are representative of at least three independent experiments.

    Article Snippet: For supershift experiments, phospho-c-Jun antibody (KM-1, sc-822X; Santa Cruz Biotechnology, Santa Cruz, CA) was preincubated on ice for 1 hour with nuclear extract, followed by the addition of the other components for 20 minutes at room temperature.

    Techniques: Binding Assay, Sequencing, Mutagenesis, Enzyme-linked Immunosorbent Assay, Activity Assay, Immunoprecipitation

    Co-immunoprecipitation assay of HNF4α and c-Jun interaction-HepG2 cell extracts treated with IL-1β (5 ng/ml) for the time indicated were immunoprecipitated with rabbit anti-HNF4α antibody as described in Materials and Methods. Immunobolot analysis was performed with goat anti-HNF4α, anti-c-Jun and anti-phospho c-Jun antibodies. Five percent of the cell lysate was immunoblotted with anti-actin antibody as internal control. Rabbit non-immune IgG was used as a negative control. The ratios of c-Jun to HNF4α and phospho-c-Jun to HNF4α are indicated below the panels.

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Bile acids and cytokines inhibit the human cholesterol 7?-hydroxylase gene via the JNK/c-Jun pathway

    doi: 10.1002/hep.21183

    Figure Lengend Snippet: Co-immunoprecipitation assay of HNF4α and c-Jun interaction-HepG2 cell extracts treated with IL-1β (5 ng/ml) for the time indicated were immunoprecipitated with rabbit anti-HNF4α antibody as described in Materials and Methods. Immunobolot analysis was performed with goat anti-HNF4α, anti-c-Jun and anti-phospho c-Jun antibodies. Five percent of the cell lysate was immunoblotted with anti-actin antibody as internal control. Rabbit non-immune IgG was used as a negative control. The ratios of c-Jun to HNF4α and phospho-c-Jun to HNF4α are indicated below the panels.

    Article Snippet: The immunoprecipitants (IP) were analyzed by immunoblot analysis using goat anti-HNF4α, anti-c-Jun or anti-phospho-c-Jun antibodies (Santa Cruz).

    Techniques: Co-Immunoprecipitation Assay, Immunoprecipitation, Negative Control

    Immunohistochemistry results for phospho-c-jun in FAP adenoma sections. Panels A and B show the typical pattern of nuclear staining for phospho-c-jun in adenomas. The table summarises the mRNA expression data for each of the nuclear phospho-c-jun-positive and -negative samples (D, down-regulation; U, up-regulation; N, no change). The relative expression values are calculated by comparing gene expression in adenoma tissue with expression in matched normal mucosa.

    Journal: British Journal of Cancer

    Article Title: Wnt signalling in adenomas of familial adenomatous polyposis patients

    doi: 10.1038/sj.bjc.6605790

    Figure Lengend Snippet: Immunohistochemistry results for phospho-c-jun in FAP adenoma sections. Panels A and B show the typical pattern of nuclear staining for phospho-c-jun in adenomas. The table summarises the mRNA expression data for each of the nuclear phospho-c-jun-positive and -negative samples (D, down-regulation; U, up-regulation; N, no change). The relative expression values are calculated by comparing gene expression in adenoma tissue with expression in matched normal mucosa.

    Article Snippet: Sections were incubated overnight with a primary antibody recognising phospho-c-jun (ser63-P; Santa Cruz Biotechnology, Heidelberg, Germany) at a dilution of 1 : 400, diluted in 20% normal goat serum.

    Techniques: Immunohistochemistry, Staining, Expressing

    Differential association of AP-1 components and ERβ with the PTPRO promoter in the presence of E2 and tamoxifen. A, Whole-cell extracts from Hs578t, 48R, and MCF-7 cells were subjected to Western blot analysis with anti-c-Fos and anti-c-Jun antibody.

    Journal: Molecular Endocrinology

    Article Title: Estrogen-Mediated Suppression of the Gene Encoding Protein Tyrosine Phosphatase PTPRO in Human Breast Cancer: Mechanism and Role in Tamoxifen Sensitivity

    doi: 10.1210/me.2008-0211

    Figure Lengend Snippet: Differential association of AP-1 components and ERβ with the PTPRO promoter in the presence of E2 and tamoxifen. A, Whole-cell extracts from Hs578t, 48R, and MCF-7 cells were subjected to Western blot analysis with anti-c-Fos and anti-c-Jun antibody.

    Article Snippet: Chromatin prepared from each treatment group was immunoprecipitated with anti-ERβ ( ) anti-c-Fos, or anti-c-Jun (both from Santa Cruz Biotechnology), following the protocol described by Ghoshal et al . ( ).

    Techniques: Western Blot

    Endothelial CYP2J2 overexpression increases PI3K/ AKT activation and altered ERK1/2 and c-Jun/JNK activation after BCCAO after BCCAO. (A) Immunoblotting (upper) of p-AKT ,total AKT and PI3K and densitometric analysis (bottom) using nuclear extract isolated from cortex 24 hours after BCCAO. p-AKT and PI3K protein upregulation after ischemia was augmented in Tie2-CYP2J2 Tr (2J2 Tr) mice, C26 inhibited the level of p-AKT and PI3K up regulated in 2J2 Tr mice. (B, C, D) Phospho-ERK1/2, ERK1/2, phosphor-c-Jun, c-Jun, phosphor-JNK and total-JNK expression in brain homogenates was assessed by immunblotting and densitometric analysis. Phospho-ERK1/2 (panels B) was significantly increased after BCCAO in Tie-CYP2J2-Tr (2J2 Tr) mice compared to WT. Phosphorylation of c-Jun (panels C) was increased after ischemia in WT but not in Tie2-CYP2J2 Tr brains. Phosphorylation of JNK (panels D) was increased after ischemia in WT but not in Tie2-CYP2J2-Tr brains. However, applied C26 reduced these effect. Blots are representative of three independent experiments. * p

    Journal: Prostaglandins & other lipid mediators

    Article Title: Cytochrome P450 2J2 Is Protective against Global Cerebral Ischemia in Transgenic Mice

    doi: 10.1016/j.prostaglandins.2012.09.004

    Figure Lengend Snippet: Endothelial CYP2J2 overexpression increases PI3K/ AKT activation and altered ERK1/2 and c-Jun/JNK activation after BCCAO after BCCAO. (A) Immunoblotting (upper) of p-AKT ,total AKT and PI3K and densitometric analysis (bottom) using nuclear extract isolated from cortex 24 hours after BCCAO. p-AKT and PI3K protein upregulation after ischemia was augmented in Tie2-CYP2J2 Tr (2J2 Tr) mice, C26 inhibited the level of p-AKT and PI3K up regulated in 2J2 Tr mice. (B, C, D) Phospho-ERK1/2, ERK1/2, phosphor-c-Jun, c-Jun, phosphor-JNK and total-JNK expression in brain homogenates was assessed by immunblotting and densitometric analysis. Phospho-ERK1/2 (panels B) was significantly increased after BCCAO in Tie-CYP2J2-Tr (2J2 Tr) mice compared to WT. Phosphorylation of c-Jun (panels C) was increased after ischemia in WT but not in Tie2-CYP2J2 Tr brains. Phosphorylation of JNK (panels D) was increased after ischemia in WT but not in Tie2-CYP2J2-Tr brains. However, applied C26 reduced these effect. Blots are representative of three independent experiments. * p

    Article Snippet: Akt, JNK, Bcl-2, Bcl-xl, Bax, c-Jun and phosphor-c-Jun were from Santa Cruz (Santa Cruz, CA).

    Techniques: Over Expression, Activation Assay, Isolation, Mouse Assay, Expressing

    TGF-β3-induced autophagy contributed to increased MUC5AC production by activating the AP1. (a-b) 16HBE cells were transduced with ATG5-siRNA lentivirus (a) and BECN1-siRNA lentivirus (b), respectively. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (c-d) 16HBE cells were treated with TGF-β3 in the presence of 3-MA (c) or Baf A1 (d). Then, the expression of phospho-c-Jun and c-Jun were detected using western blot assay. (e) 16HBE cells were transfected with Smad2/3-siRNA. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (f) 16HBE cells were transfected with c-Jun-siRNA, after treating with TGF-β3 (10 ng/ml) for 24 h, and then phospho-c-Jun and c-Jun were detected by western blot. (g) Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells were transfected with c-Jun-siRNA. (h) Quantitation of fluorescence intensity of MUC5AC (each group n = 10 images for quantification). (i) 16HBE cells were transfected with c-Jun-siRNA. Real-time PCR was performed to detect the expression of MUC5AC gene after treated with TGF-β3 (10 ng/ml). Data are representative of the three independent experiments and are presented as means ± s.d . *P

    Journal: EBioMedicine

    Article Title: TGF-β3 Promotes MUC5AC Hyper-Expression by Modulating Autophagy Pathway in Airway Epithelium

    doi: 10.1016/j.ebiom.2018.06.032

    Figure Lengend Snippet: TGF-β3-induced autophagy contributed to increased MUC5AC production by activating the AP1. (a-b) 16HBE cells were transduced with ATG5-siRNA lentivirus (a) and BECN1-siRNA lentivirus (b), respectively. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (c-d) 16HBE cells were treated with TGF-β3 in the presence of 3-MA (c) or Baf A1 (d). Then, the expression of phospho-c-Jun and c-Jun were detected using western blot assay. (e) 16HBE cells were transfected with Smad2/3-siRNA. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, phospho-c-Jun and c-Jun were detected by western blot. (f) 16HBE cells were transfected with c-Jun-siRNA, after treating with TGF-β3 (10 ng/ml) for 24 h, and then phospho-c-Jun and c-Jun were detected by western blot. (g) Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells were transfected with c-Jun-siRNA. (h) Quantitation of fluorescence intensity of MUC5AC (each group n = 10 images for quantification). (i) 16HBE cells were transfected with c-Jun-siRNA. Real-time PCR was performed to detect the expression of MUC5AC gene after treated with TGF-β3 (10 ng/ml). Data are representative of the three independent experiments and are presented as means ± s.d . *P

    Article Snippet: The control siRNA, Smad2/3 siRNA and c-Jun siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Transduction, Western Blot, Expressing, Transfection, Immunofluorescence, Quantitation Assay, Fluorescence, Real-time Polymerase Chain Reaction

    Smad2/3 pathway is involved in TGF-β3 induced autophagy and MUC5AC. (a) 16HBE cells were transfected with Smad2/3-siRNA. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, LC3B, BECN1, ATG5, phospho-Smad2, Smad2, phospho-Smad3 and Smad3 were detected by western blot. (b) Relative changes in the density of LC3B II were detected. (c) 16HBE cells that stably expressed mCherry-EGFP-LC3 fusion protein were transfected with Smad2/3-siRNA. After treating with TGF-β3 (10 ng/ml) for 24 h, autophagosomes were observed under confocal microscope (2000× magnification) in 16HBE cells. Bar scale, 5 mm. (d) Quantification of the number of LC3 puncta (each group n = 10 images for quantification). (e) 16HBE cells were transfected with Smad2/3-siRNA. Real-time PCR was performed to detect the expression of MUC5AC gene after treated with TGF-β3 (10 ng/ml). (f) Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells transfected with Smad2/3-siRNA. (g) Quantitation of the fluorescence intensity of MUC5AC (each group n = 10 images for quantification). Data are representative of three independent experiments and are presented as means ± s.d . *P

    Journal: EBioMedicine

    Article Title: TGF-β3 Promotes MUC5AC Hyper-Expression by Modulating Autophagy Pathway in Airway Epithelium

    doi: 10.1016/j.ebiom.2018.06.032

    Figure Lengend Snippet: Smad2/3 pathway is involved in TGF-β3 induced autophagy and MUC5AC. (a) 16HBE cells were transfected with Smad2/3-siRNA. After treating the cells with TGF-β3 (10 ng/ml) for 24 h, LC3B, BECN1, ATG5, phospho-Smad2, Smad2, phospho-Smad3 and Smad3 were detected by western blot. (b) Relative changes in the density of LC3B II were detected. (c) 16HBE cells that stably expressed mCherry-EGFP-LC3 fusion protein were transfected with Smad2/3-siRNA. After treating with TGF-β3 (10 ng/ml) for 24 h, autophagosomes were observed under confocal microscope (2000× magnification) in 16HBE cells. Bar scale, 5 mm. (d) Quantification of the number of LC3 puncta (each group n = 10 images for quantification). (e) 16HBE cells were transfected with Smad2/3-siRNA. Real-time PCR was performed to detect the expression of MUC5AC gene after treated with TGF-β3 (10 ng/ml). (f) Representative immunofluorescence images of TGF-β3-induced MUC5AC in 16HBE cells transfected with Smad2/3-siRNA. (g) Quantitation of the fluorescence intensity of MUC5AC (each group n = 10 images for quantification). Data are representative of three independent experiments and are presented as means ± s.d . *P

    Article Snippet: The control siRNA, Smad2/3 siRNA and c-Jun siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Transfection, Western Blot, Stable Transfection, Microscopy, Real-time Polymerase Chain Reaction, Expressing, Immunofluorescence, Quantitation Assay, Fluorescence

    Recombinant full-length dimeric c-Jun-containing human AP-1 complexes are all active in binding the consensus TRE sequence found in the human cyclin D1 gene. A , shown is a schematic drawing of human c-Jun and Fos family proteins tagged at the N terminus

    Journal: The Journal of Biological Chemistry

    Article Title: Binding Site Specificity and Factor Redundancy in Activator Protein-1-driven Human Papillomavirus Chromatin-dependent Transcription *

    doi: 10.1074/jbc.M111.290874

    Figure Lengend Snippet: Recombinant full-length dimeric c-Jun-containing human AP-1 complexes are all active in binding the consensus TRE sequence found in the human cyclin D1 gene. A , shown is a schematic drawing of human c-Jun and Fos family proteins tagged at the N terminus

    Article Snippet: For antibody supershift assay, 68 or 200 ng of rabbit polyclonal antibodies against the hexahistidine tag (sc-804, Santa Cruz Biotechnology) or the N-terminal 79 amino acids of human c-Jun (sc-1694, Santa Cruz Biotechnology) were added 10 min before the termination of the reaction.

    Techniques: Recombinant, Binding Assay, Sequencing

    Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with polyclonal c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal anti-c-Jun antibodies (upper panel) or anti-Pin1 antibodies (lower panel).

    Journal: The EMBO Journal

    Article Title: Pin1 is overexpressed in breast cancer and cooperates with Ras signaling in increasing the transcriptional activity of c-Jun towards cyclin D1

    doi: 10.1093/emboj/20.13.3459

    Figure Lengend Snippet: Fig. 4. Pin1 binds to c-Jun phosphorylated on Ser 63/73 -Pro. ( A and B ) Modulation of c-Jun phosphorylation by Ras or JNK. HeLa cells were co-transfected with c-Jun or c-Jun S63/73A and Ha-Ras, DN-Ras, activated JNK or control vector. Cells were harvested and cellular proteins were subjected to immunoblotting analysis with antibodies against c-Jun (A) or phosphorylated Ser 63/73 -c-Jun (B). ( C and D ) Interaction between Pin1 and c-Jun phosphorylated on Ser 63/73 -Pro. The same cellular proteins as those described in (A) were incubated with GST–agarose beads that had been pre-incubated with either GST alone or GST–Pin1. Proteins associated with the beads were subjected to immunoblotting analysis with antibodies against c-Jun (C) or phosphorylated Ser 63/73 ). ( E and F ) No interaction between Pin1 and c-Jun S63/73A . The same cellular proteins as those described in the (A) were incubated with GST–agarose beads containing GST or GST–Pin1, and bound proteins were subjected to immunoblotting analysis with antibodies against c-Jun (E) or phosphorylated Ser 63/73 -c-Jun (F). ( G and H ) Co-immunoprecipitation of transfected (G) or endogenous (H) c-Jun with endogenous Pin1. HeLa cells were co-transfected with c-Jun and Ha-Ras or JNK. c-Jun was immunoprecipitated from transfected HeLa cells (G) or non-transfected breast cancer cell lines (H) with polyclonal c-Jun antibodies or non-related antibodies (Control), and then subjected to immunoblotting using monoclonal anti-c-Jun antibodies (upper panel) or anti-Pin1 antibodies (lower panel).

    Article Snippet: For co-immunoprecipitation, we used anti-c-Jun polyclonal antibodies (Santa Cruz) and unrelated polyclonal antibodies (Pericentrin antibodies) as a control.

    Techniques: Transfection, Plasmid Preparation, Incubation, Immunoprecipitation

    C-Jun was required for TNC-induced MMP9 expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and anti-c-Jun (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P

    Journal: Oncotarget

    Article Title: Tenascin-C induces migration and invasion through JNK/c-Jun signalling in pancreatic cancer

    doi: 10.18632/oncotarget.20160

    Figure Lengend Snippet: C-Jun was required for TNC-induced MMP9 expression (A) The sequence and position of the c-Jun binding site in the MMP9 promoter are shown. (B) ChIP of c-Jun on the promoter of MMP9 . PANC-1 cells were transfected with empty siCtrl, siTNC, vector or TNC plasmid. PCR amplification from the total chromatin (bottom) was used as a positive control, anti-IgG (middle) served as a negative control, and anti-c-Jun (top) showed the interaction between the c-Jun and MMP9 promoter after the indicated treatment. (C) TNC transactivates the MMP9 promoter via c-Jun. The luciferase activity of the reporters in the indicated cells was assessed by a dual-luciferase reporter assay. The relative luciferase activity is the ratio of the luciferase activity in each of the tested cells to that in the control cells. Data represent the mean ± SD. (n = 3, * P

    Article Snippet: The primary antibodies were rabbit anti- E-cadherin, rabbit anti-N-cadherin, rabbit anti-Vimentin (Cell Signaling Technology, Danvers, MA, USA), mouse anti-MMP9, mouse anti-MMP2, mouse anti-JNK1, mouse anti-p-JNK, rabbit anti-c-Jun, mouse anti-p-c-Jun, mouse anti-FAK, mouse anti-β-actin (Santa Cruz Biotechnology), rabbit anti-Paxillin, and rabbit anti-p-Paxillin (Abcam, Cambridge, MA, USA).

    Techniques: Expressing, Sequencing, Binding Assay, Chromatin Immunoprecipitation, Transfection, Plasmid Preparation, Polymerase Chain Reaction, Amplification, Positive Control, Negative Control, Luciferase, Activity Assay, Reporter Assay

    ILK-induced AP-1 activity is mediated by c-jun. (A) In this band shift assay, serum-exposed HEK-293 cells were transfected by Lipofectin with 0.5 μg of each of the indicated cDNAs. PD98059 (25 μM) was added to the medium 12 h prior to harvesting the cells. Nuclear extracts (2 μg) from the transfected cells were incubated with 32 P-end-labeled AP-1 consensus oligonucleotide containing the protein binding site. Reaction products were analyzed on a nondenaturing 5% polyacrylamide gel (left gel). The specificity of complex formation was established by a competition experiment using cold AP-1 oligonucleotide as the competitor (right gel). For immunoblot studies, 10 μg of protein was resolved by SDS–10% PAGE. c-jun (Ser-73) phosphorylation, c-jun protein expression level, ERK phosphorylation (Tyr-204), and ERK protein expression level (not shown) were determined by Western blot analysis. ILK and GSK-3 expression levels in the transfected cells were evaluated by Western blot analysis using anti-V5 and anti-HA antibodies, respectively. (B) Anti-c-jun antibody shifts ILK-induced AP-1 complex. Nuclear extracts (2 μg) from HEK-293 cells transfected with ILK-V5 cDNA were incubated with 32 P-AP-1 oligonucleotide in the absence or presence of anti-c-jun antibody or nonspecific IgG (10 μg). The latter did not induce a mobility shift of the complex (not shown).

    Journal: Molecular and Cellular Biology

    Article Title: Cell-Extracellular Matrix Interactions Stimulate the AP-1 Transcription Factor in an Integrin-Linked Kinase- and Glycogen Synthase Kinase 3-Dependent Manner

    doi:

    Figure Lengend Snippet: ILK-induced AP-1 activity is mediated by c-jun. (A) In this band shift assay, serum-exposed HEK-293 cells were transfected by Lipofectin with 0.5 μg of each of the indicated cDNAs. PD98059 (25 μM) was added to the medium 12 h prior to harvesting the cells. Nuclear extracts (2 μg) from the transfected cells were incubated with 32 P-end-labeled AP-1 consensus oligonucleotide containing the protein binding site. Reaction products were analyzed on a nondenaturing 5% polyacrylamide gel (left gel). The specificity of complex formation was established by a competition experiment using cold AP-1 oligonucleotide as the competitor (right gel). For immunoblot studies, 10 μg of protein was resolved by SDS–10% PAGE. c-jun (Ser-73) phosphorylation, c-jun protein expression level, ERK phosphorylation (Tyr-204), and ERK protein expression level (not shown) were determined by Western blot analysis. ILK and GSK-3 expression levels in the transfected cells were evaluated by Western blot analysis using anti-V5 and anti-HA antibodies, respectively. (B) Anti-c-jun antibody shifts ILK-induced AP-1 complex. Nuclear extracts (2 μg) from HEK-293 cells transfected with ILK-V5 cDNA were incubated with 32 P-AP-1 oligonucleotide in the absence or presence of anti-c-jun antibody or nonspecific IgG (10 μg). The latter did not induce a mobility shift of the complex (not shown).

    Article Snippet: For the supershift assay, 10 μg of rabbit anti-c-jun antibody (Santa Cruz Biotechnology) or nonspecific IgG was added to the reaction mixture, subsequent to the addition of the 32 P-labeled oligonucleotide probe, and the mixture was incubated for 45 min at room temperature.

    Techniques: Activity Assay, Electrophoretic Mobility Shift Assay, Transfection, Incubation, Labeling, Protein Binding, Polyacrylamide Gel Electrophoresis, Expressing, Western Blot, Mobility Shift