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  • 99
    Thermo Fisher isopropyl thiogalactoside iptg
    Detection and purification of expressed OipA. (A) OipA was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Lane M: low molecular weight standard protein size marker (kDa); lane 1: non-induced OipA; lane 2, 3, 4, and 5: induced OipA with 1 <t>mmol/L</t> <t>IPTG</t> at t = 0, 1, 2, and 3 hr, respectively. 30-kD band were detected in induced lanes. T = 3 hr was the best time for induction. (B) Purified OipA was detected using SDS-PAGE. Lane M: low molecular weight standard protein size marker (kDa); lane 1: 30-kD band was detected as purified OipA.
    Isopropyl Thiogalactoside Iptg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 24 article reviews
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    99
    Millipore iptg
    Optimization of a XNOR circuit with the sRNA pool. ( A ) The circuit diagram for the XNOR function is shown. Inside each NOR gate, the repressor name is shown. ( B ) Truth table for a two-input XNOR gate. ( C ) The genetic design for the XNOR circuit is shown. Blunt ended lines represent the repression of promoters. The plasmid map and part sequences are provided in Supplementary Figure S1 and Table S3 . ( D ) Response of an untuned XNOR circuit where empty vector is used in place of a sRNA pool plasmid. Inputs correspond to the absence or presence of 1 mM <t>IPTG</t> (left ‘0’ / ‘1’) and <t>aTC</t> (2 ng/ml; right ‘0’ / ‘1’). ( E ) Sort gates (shown in gray) used for each input combination are shown and numbered in the order in which they were applied to the population. The sorting procedure is described in Materials and Methods. ( F ) Response of the best sRNA-tuned XNOR circuit. ( G ) The sRNA-mediated knockdown of each repressor gene in the tuned circuit. ( H ) Calculated RBS strengths for the original untuned circuit (black bars) and for the reconstructed (new RBSs) tuned circuit (white bars) (Materials and Methods). ( I ) Response of the circuit with the substituted RBSs. For all data, error bars correspond to the standard deviation of three experiments performed on different days.
    Iptg, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9488 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Research Products International isopropyl thiogalactoside iptg
    Optimization of a XNOR circuit with the sRNA pool. ( A ) The circuit diagram for the XNOR function is shown. Inside each NOR gate, the repressor name is shown. ( B ) Truth table for a two-input XNOR gate. ( C ) The genetic design for the XNOR circuit is shown. Blunt ended lines represent the repression of promoters. The plasmid map and part sequences are provided in Supplementary Figure S1 and Table S3 . ( D ) Response of an untuned XNOR circuit where empty vector is used in place of a sRNA pool plasmid. Inputs correspond to the absence or presence of 1 mM <t>IPTG</t> (left ‘0’ / ‘1’) and <t>aTC</t> (2 ng/ml; right ‘0’ / ‘1’). ( E ) Sort gates (shown in gray) used for each input combination are shown and numbered in the order in which they were applied to the population. The sorting procedure is described in Materials and Methods. ( F ) Response of the best sRNA-tuned XNOR circuit. ( G ) The sRNA-mediated knockdown of each repressor gene in the tuned circuit. ( H ) Calculated RBS strengths for the original untuned circuit (black bars) and for the reconstructed (new RBSs) tuned circuit (white bars) (Materials and Methods). ( I ) Response of the circuit with the substituted RBSs. For all data, error bars correspond to the standard deviation of three experiments performed on different days.
    Isopropyl Thiogalactoside Iptg, supplied by Research Products International, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GE Healthcare isopropyl thiogalactoside iptg
    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of recombinant interleukin-6 (rIL-6) ( A ) and recombinant outer membrane protein V (rOmpV) ( B ). Lane M: protein marker; Lanes 1and 4: recombinant clones without isopropy-β- d <t>-thiogalactoside</t> <t>(IPTG)</t> induction; Lanes 2 and 5: recombinant clones with IPTG induction; Lanes 3 and 6: purified rIL-6 and rOmpV.
    Isopropyl Thiogalactoside Iptg, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Denville Scientific iptg
    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of recombinant interleukin-6 (rIL-6) ( A ) and recombinant outer membrane protein V (rOmpV) ( B ). Lane M: protein marker; Lanes 1and 4: recombinant clones without isopropy-β- d <t>-thiogalactoside</t> <t>(IPTG)</t> induction; Lanes 2 and 5: recombinant clones with IPTG induction; Lanes 3 and 6: purified rIL-6 and rOmpV.
    Iptg, supplied by Denville Scientific, used in various techniques. Bioz Stars score: 92/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Promega iptg
    Expression and purification of recombinant Cathepsin L-like protein. Protein samples were separated by 12.5% SDS-PAGE gel electrophoresis. ( A ) Molecular weight standard, ( B ) lysate of culture before and ( C ) after induction with <t>IPTG</t> and ( D ) recombinant Cathepsin L-like protein (MW 47.6 KDa) purified by gel filtration.
    Iptg, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 638 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iptg  (TaKaRa)
    99
    TaKaRa iptg
    3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated <t>Gp78.</t> (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by <t>IPTG.</t> Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).
    Iptg, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 722 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iptg  (Abcam)
    99
    Abcam iptg
    3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated <t>Gp78.</t> (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by <t>IPTG.</t> Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).
    Iptg, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Millipore β isopropyl thiogalactoside
    3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated <t>Gp78.</t> (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by <t>IPTG.</t> Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).
    β Isopropyl Thiogalactoside, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher iptg xgal
    3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated <t>Gp78.</t> (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by <t>IPTG.</t> Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).
    Iptg Xgal, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore 1 thiogalactopyranoside iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    1 Thiogalactopyranoside Iptg, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 522 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Anatrace iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Anatrace, used in various techniques. Bioz Stars score: 93/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Avantor iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Avantor, used in various techniques. Bioz Stars score: 94/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Beijing CWBio iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Beijing CWBio, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Carl Roth GmbH iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 93/100, based on 213 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Corning Life Sciences iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Diagnostic Chemicals Ltd iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Diagnostic Chemicals Ltd, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Duchefa iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Duchefa, used in various techniques. Bioz Stars score: 95/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Euromedex iptg
    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with <t>IPTG</t> for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl <t>β-D-1-thiogalactopyranoside.</t>
    Iptg, supplied by Euromedex, used in various techniques. Bioz Stars score: 94/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Fisher Scientific iptg
    SOD activities from S. aureus sod mutants. Lane 1, RN6390; lane 2, <t>sodM</t> mutant; lane 3, sodA mutant; lane 4, double ( sodM sodA ) mutant; lane 5, double mutant containing pCL15 sodM induced with <t>IPTG;</t> lane 6, double mutant containing pCL15 sodM uninduced.
    Iptg, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 99/100, based on 349 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    FUJIFILM iptg
    Analysis of recombinant <t>GST-muCCR5-NH2</t> terminal fusion protein. Mr: MW marker, 1, 2: products purified by glutathione sepharose, 3, 4: products not purified, 1, 3: expression products induced by <t>IPTG</t>
    Iptg, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 95/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Analysis of recombinant <t>GST-muCCR5-NH2</t> terminal fusion protein. Mr: MW marker, 1, 2: products purified by glutathione sepharose, 3, 4: products not purified, 1, 3: expression products induced by <t>IPTG</t>
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    Analysis of recombinant <t>GST-muCCR5-NH2</t> terminal fusion protein. Mr: MW marker, 1, 2: products purified by glutathione sepharose, 3, 4: products not purified, 1, 3: expression products induced by <t>IPTG</t>
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    Induction of <t>p53</t> down regulates Bcl-3 protein levels in H1299 w/tp53 cells. (A) p53 represses Bcl-3 but not p100/p52 protein levels. Following 16 h of p53 induction by increasing concentrations of <t>IPTG</t> in H1299 w/tp53 cells, whole-cell lysates were prepared and the protein levels of p100/p52, Bcl-3, and β-actin control were analyzed by Western blot. (B) Bcl-3 down regulation is evident 12 h after IPTG induction of p53. p53 was induced in H1299 w/tp53 cells by 100 μM IPTG. Cells were harvested at the indicated times following IPTG treatment, and nuclear protein extracts were prepared and analyzed by Western blot for Bcl-3 levels. (C) Induction of p53 in H1299 w/tp53 cells does not repress Bcl-3 mRNA levels. RT-PCR analysis was performed by using two sets of primers specific to Bcl-3 (P1 and P2) or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.
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    Nacalai iptg
    Induction of <t>p53</t> down regulates Bcl-3 protein levels in H1299 w/tp53 cells. (A) p53 represses Bcl-3 but not p100/p52 protein levels. Following 16 h of p53 induction by increasing concentrations of <t>IPTG</t> in H1299 w/tp53 cells, whole-cell lysates were prepared and the protein levels of p100/p52, Bcl-3, and β-actin control were analyzed by Western blot. (B) Bcl-3 down regulation is evident 12 h after IPTG induction of p53. p53 was induced in H1299 w/tp53 cells by 100 μM IPTG. Cells were harvested at the indicated times following IPTG treatment, and nuclear protein extracts were prepared and analyzed by Western blot for Bcl-3 levels. (C) Induction of p53 in H1299 w/tp53 cells does not repress Bcl-3 mRNA levels. RT-PCR analysis was performed by using two sets of primers specific to Bcl-3 (P1 and P2) or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.
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    iptg  (Roche)
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    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM <t>IPTG</t> on <t>Mueller–Hinton</t> broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).
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    Valiant iptg
    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM <t>IPTG</t> on <t>Mueller–Hinton</t> broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).
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    Amresco iptg
    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM <t>IPTG</t> on <t>Mueller–Hinton</t> broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).
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    Applichem iptg
    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM <t>IPTG</t> on <t>Mueller–Hinton</t> broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).
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    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM <t>IPTG</t> on <t>Mueller–Hinton</t> broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).
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    Image Search Results


    Detection and purification of expressed OipA. (A) OipA was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Lane M: low molecular weight standard protein size marker (kDa); lane 1: non-induced OipA; lane 2, 3, 4, and 5: induced OipA with 1 mmol/L IPTG at t = 0, 1, 2, and 3 hr, respectively. 30-kD band were detected in induced lanes. T = 3 hr was the best time for induction. (B) Purified OipA was detected using SDS-PAGE. Lane M: low molecular weight standard protein size marker (kDa); lane 1: 30-kD band was detected as purified OipA.

    Journal: Clinical and Experimental Vaccine Research

    Article Title: Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

    doi: 10.7774/cevr.2015.4.2.177

    Figure Lengend Snippet: Detection and purification of expressed OipA. (A) OipA was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Lane M: low molecular weight standard protein size marker (kDa); lane 1: non-induced OipA; lane 2, 3, 4, and 5: induced OipA with 1 mmol/L IPTG at t = 0, 1, 2, and 3 hr, respectively. 30-kD band were detected in induced lanes. T = 3 hr was the best time for induction. (B) Purified OipA was detected using SDS-PAGE. Lane M: low molecular weight standard protein size marker (kDa); lane 1: 30-kD band was detected as purified OipA.

    Article Snippet: Expression of recombinant protein was induced with 1 mmol/L of IPTG (Thermo Scientific, Waltham, MA, USA).

    Techniques: Purification, Polyacrylamide Gel Electrophoresis, SDS Page, Molecular Weight, Marker

    Optimization of a XNOR circuit with the sRNA pool. ( A ) The circuit diagram for the XNOR function is shown. Inside each NOR gate, the repressor name is shown. ( B ) Truth table for a two-input XNOR gate. ( C ) The genetic design for the XNOR circuit is shown. Blunt ended lines represent the repression of promoters. The plasmid map and part sequences are provided in Supplementary Figure S1 and Table S3 . ( D ) Response of an untuned XNOR circuit where empty vector is used in place of a sRNA pool plasmid. Inputs correspond to the absence or presence of 1 mM IPTG (left ‘0’ / ‘1’) and aTC (2 ng/ml; right ‘0’ / ‘1’). ( E ) Sort gates (shown in gray) used for each input combination are shown and numbered in the order in which they were applied to the population. The sorting procedure is described in Materials and Methods. ( F ) Response of the best sRNA-tuned XNOR circuit. ( G ) The sRNA-mediated knockdown of each repressor gene in the tuned circuit. ( H ) Calculated RBS strengths for the original untuned circuit (black bars) and for the reconstructed (new RBSs) tuned circuit (white bars) (Materials and Methods). ( I ) Response of the circuit with the substituted RBSs. For all data, error bars correspond to the standard deviation of three experiments performed on different days.

    Journal: Nucleic Acids Research

    Article Title: Balancing gene expression without library construction via a reusable sRNA pool

    doi: 10.1093/nar/gkx530

    Figure Lengend Snippet: Optimization of a XNOR circuit with the sRNA pool. ( A ) The circuit diagram for the XNOR function is shown. Inside each NOR gate, the repressor name is shown. ( B ) Truth table for a two-input XNOR gate. ( C ) The genetic design for the XNOR circuit is shown. Blunt ended lines represent the repression of promoters. The plasmid map and part sequences are provided in Supplementary Figure S1 and Table S3 . ( D ) Response of an untuned XNOR circuit where empty vector is used in place of a sRNA pool plasmid. Inputs correspond to the absence or presence of 1 mM IPTG (left ‘0’ / ‘1’) and aTC (2 ng/ml; right ‘0’ / ‘1’). ( E ) Sort gates (shown in gray) used for each input combination are shown and numbered in the order in which they were applied to the population. The sorting procedure is described in Materials and Methods. ( F ) Response of the best sRNA-tuned XNOR circuit. ( G ) The sRNA-mediated knockdown of each repressor gene in the tuned circuit. ( H ) Calculated RBS strengths for the original untuned circuit (black bars) and for the reconstructed (new RBSs) tuned circuit (white bars) (Materials and Methods). ( I ) Response of the circuit with the substituted RBSs. For all data, error bars correspond to the standard deviation of three experiments performed on different days.

    Article Snippet: IPTG (Sigma-Aldrich, I6758) and aTC (37919) were used as inducers.

    Techniques: Plasmid Preparation, Standard Deviation

    Expression dynamics of SGIV VP19. (A) SDS-PAGE analysis of purified recombinant SGIV VP19. Lines M, 1, 2, 3, 4 and 5 showed protein markers, pET-VP19 (uninduced), pET-VP19 (IPTG induced), supernatant, pellet of induced pET-VP19 and the purified pET-VP19, respectively. (B) The specificity of anti-VP19 serum. SGIV or mock infected cells at 48 h p.i. were collected and centrifuged for SDS-PAGE and western blotting. Line M, 1 and 2 showed protein markers, mock and SGIV infected cells at 48 h p.i., respectively. (C) The expression pattern of VP19 during SGIV infection. Actin was chosen as the internal control. (D) VP19 was identified as a late protein. Cells were infected with SGIV at MOI of 0.5 for 6 h and 24 h under the treatment with CHX or AraC, respectively. Then cells were collected for western blotting analysis. The late protein MCP was chosen as a positive control.

    Journal: Virology Journal

    Article Title: Characterization of an envelope gene VP19 from Singapore grouper iridovirus

    doi: 10.1186/1743-422X-10-354

    Figure Lengend Snippet: Expression dynamics of SGIV VP19. (A) SDS-PAGE analysis of purified recombinant SGIV VP19. Lines M, 1, 2, 3, 4 and 5 showed protein markers, pET-VP19 (uninduced), pET-VP19 (IPTG induced), supernatant, pellet of induced pET-VP19 and the purified pET-VP19, respectively. (B) The specificity of anti-VP19 serum. SGIV or mock infected cells at 48 h p.i. were collected and centrifuged for SDS-PAGE and western blotting. Line M, 1 and 2 showed protein markers, mock and SGIV infected cells at 48 h p.i., respectively. (C) The expression pattern of VP19 during SGIV infection. Actin was chosen as the internal control. (D) VP19 was identified as a late protein. Cells were infected with SGIV at MOI of 0.5 for 6 h and 24 h under the treatment with CHX or AraC, respectively. Then cells were collected for western blotting analysis. The late protein MCP was chosen as a positive control.

    Article Snippet: After IPTG induction of E. coli BL21 (DE3) containing pET-VP19t, the recombinant fusion protein rVP19t was purified using the HisBind purification kit (Novagen, Germany) according to the manufacture’s protocol.

    Techniques: Expressing, SDS Page, Purification, Recombinant, Positron Emission Tomography, Infection, Western Blot, Positive Control

    Complementation of a proBA mutation by tomPRO1 and tomPRO2 , and their products expressed in E. coli . A and B, Expression vectors containing the tomPRO1 and tomPRO2 cDNA clones were introduced into strain KC1325 (a derivative of BL21[DL3]pLysS carrying the proB1658 :: Tn10 insertion, which is polar on proA ). a, KC1325 harboring the vector, pKS only. b, KC1325 harboring pPRO1. c, KC1325 harboring pET32a only. d, KC1325 harboring pET32PRO2. Strain KC1325 containing each plasmid was streaked on minimal M63 medium containing Glc, thiamine, and IPTG with (A) and without (B) Pro, and incubated for 2 d at 37°C. All strains could grow on the media supplemented with Pro (A). C, Total cell extracts from either E. coli strain HB101, containing pKS (lane 1) and pPRO1 (lane 2), or strain KC1325, containing pET32a (lane 3) and pET32PRO2 (lane 4), were analyzed by SDS-PAGE. The gels were stained with Coomasie brilliant blue. tomPRO1 products are indicated as GK and GPR, and the tomPRO2 product as P5CS. Numbers at left refer to size standards (in kD).

    Journal: Plant Physiology

    Article Title: Comparative Analysis of the Regulation of Expression and Structures of Two Evolutionarily Divergent Genes for ?1-Pyrroline-5-Carboxylate Synthetase from Tomato 1

    doi:

    Figure Lengend Snippet: Complementation of a proBA mutation by tomPRO1 and tomPRO2 , and their products expressed in E. coli . A and B, Expression vectors containing the tomPRO1 and tomPRO2 cDNA clones were introduced into strain KC1325 (a derivative of BL21[DL3]pLysS carrying the proB1658 :: Tn10 insertion, which is polar on proA ). a, KC1325 harboring the vector, pKS only. b, KC1325 harboring pPRO1. c, KC1325 harboring pET32a only. d, KC1325 harboring pET32PRO2. Strain KC1325 containing each plasmid was streaked on minimal M63 medium containing Glc, thiamine, and IPTG with (A) and without (B) Pro, and incubated for 2 d at 37°C. All strains could grow on the media supplemented with Pro (A). C, Total cell extracts from either E. coli strain HB101, containing pKS (lane 1) and pPRO1 (lane 2), or strain KC1325, containing pET32a (lane 3) and pET32PRO2 (lane 4), were analyzed by SDS-PAGE. The gels were stained with Coomasie brilliant blue. tomPRO1 products are indicated as GK and GPR, and the tomPRO2 product as P5CS. Numbers at left refer to size standards (in kD).

    Article Snippet: Production of a recombinant protein for tomPRO2 was induced by 1 m m IPTG at 25°C for 17 h, based on the manufacturer's instructions (Novagen).

    Techniques: Mutagenesis, Expressing, Clone Assay, Plasmid Preparation, Gas Chromatography, Incubation, SDS Page, Staining

    Recruitment of HJURP to the LacO array by LacI-tagged CENP-A is IPTG sensitive, but recruitment of CENP-C is IPTG insensitive. (A) Experimental scheme for this study. (B and D) Quantitation of HJURP intensity (B) and CENP-C intensity (D) at the LacO array normalized to the mean intensity at endogenous centromeres for HA-LacI–tagged CENP-A ±IPTG treatment. (C and E) Representative images of HJURP (C) or CENP-C (E) recruitment to the LacO array by HA-LacI–tagged CENP-A ±IPTG treatment. (F) Quantitation of CENP-C intensity at the LacO array 24 and 48 h after transfection of HA-LacI–tagged CENP-A. (G) Representative images of CENP-C recruitment to the LacO array 24 and 48 h after transfection of HA-LacI–tagged CENP-A. (H) Diagram of chimeras used in I–L. (I) Quantitation of HJURP intensity at the LacO array for the indicated chimeras. (J) Representative images of HJURP recruitment to the LacO array by the indicated chimeras fused to HA-LacI. (K) Quantitation of CENP-A intensity at the LacO array for the indicated chimeras. Error bars show SEM. (L) Representative images of CENP-A recruitment to the LacO array by the indicated chimeras. Bars: (main images) 5 µm; (insets) 1 µm. Insets show magnification of the boxed regions. For all graphs, an asterisk denotes significant differences (*, P

    Journal: The Journal of Cell Biology

    Article Title: Both tails and the centromere targeting domain of CENP-A are required for centromere establishment

    doi: 10.1083/jcb.201412011

    Figure Lengend Snippet: Recruitment of HJURP to the LacO array by LacI-tagged CENP-A is IPTG sensitive, but recruitment of CENP-C is IPTG insensitive. (A) Experimental scheme for this study. (B and D) Quantitation of HJURP intensity (B) and CENP-C intensity (D) at the LacO array normalized to the mean intensity at endogenous centromeres for HA-LacI–tagged CENP-A ±IPTG treatment. (C and E) Representative images of HJURP (C) or CENP-C (E) recruitment to the LacO array by HA-LacI–tagged CENP-A ±IPTG treatment. (F) Quantitation of CENP-C intensity at the LacO array 24 and 48 h after transfection of HA-LacI–tagged CENP-A. (G) Representative images of CENP-C recruitment to the LacO array 24 and 48 h after transfection of HA-LacI–tagged CENP-A. (H) Diagram of chimeras used in I–L. (I) Quantitation of HJURP intensity at the LacO array for the indicated chimeras. (J) Representative images of HJURP recruitment to the LacO array by the indicated chimeras fused to HA-LacI. (K) Quantitation of CENP-A intensity at the LacO array for the indicated chimeras. Error bars show SEM. (L) Representative images of CENP-A recruitment to the LacO array by the indicated chimeras. Bars: (main images) 5 µm; (insets) 1 µm. Insets show magnification of the boxed regions. For all graphs, an asterisk denotes significant differences (*, P

    Article Snippet: For U20S experiments involving IPTG treatment, 48 h after transfection, cells were treated or untreated with 15 mM IPTG (Sigma-Aldrich) for 1 h and processed for indirect immunofluorescence, as described earlier in this section.

    Techniques: Quantitation Assay, Transfection

    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of recombinant interleukin-6 (rIL-6) ( A ) and recombinant outer membrane protein V (rOmpV) ( B ). Lane M: protein marker; Lanes 1and 4: recombinant clones without isopropy-β- d -thiogalactoside (IPTG) induction; Lanes 2 and 5: recombinant clones with IPTG induction; Lanes 3 and 6: purified rIL-6 and rOmpV.

    Journal: International Journal of Molecular Sciences

    Article Title: The Immune Adjuvant Effects of Flounder (Paralichthys olivaceus) Interleukin-6 on E. tarda Subunit Vaccine OmpV

    doi: 10.3390/ijms18071445

    Figure Lengend Snippet: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of recombinant interleukin-6 (rIL-6) ( A ) and recombinant outer membrane protein V (rOmpV) ( B ). Lane M: protein marker; Lanes 1and 4: recombinant clones without isopropy-β- d -thiogalactoside (IPTG) induction; Lanes 2 and 5: recombinant clones with IPTG induction; Lanes 3 and 6: purified rIL-6 and rOmpV.

    Article Snippet: When the OD600 of the cultures reached 0.6, 1 mM isopropyl thiogalactoside (IPTG) was added and grown at 37 °C for an additional 10 h. Then, the cultures were centrifuged, and the His-tagged rIL-6 was purified using His TrapTM HP Ni-Agarose (GE healthcare, Beijing, China).

    Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Recombinant, Marker, Clone Assay, Purification

    Expression and purification of recombinant Cathepsin L-like protein. Protein samples were separated by 12.5% SDS-PAGE gel electrophoresis. ( A ) Molecular weight standard, ( B ) lysate of culture before and ( C ) after induction with IPTG and ( D ) recombinant Cathepsin L-like protein (MW 47.6 KDa) purified by gel filtration.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Improving Serodiagnosis of Human and Canine Leishmaniasis with Recombinant Leishmania braziliensis Cathepsin L-like Protein and a Synthetic Peptide Containing Its Linear B-cell Epitope

    doi: 10.1371/journal.pntd.0003426

    Figure Lengend Snippet: Expression and purification of recombinant Cathepsin L-like protein. Protein samples were separated by 12.5% SDS-PAGE gel electrophoresis. ( A ) Molecular weight standard, ( B ) lysate of culture before and ( C ) after induction with IPTG and ( D ) recombinant Cathepsin L-like protein (MW 47.6 KDa) purified by gel filtration.

    Article Snippet: The recombinant CatL (r CATL) expression was performed by adding 1.0 mM IPTG (Isopropyl-β-D-thiogalactopyranoside, Promega, Canada) for 24 h at 12°C with shaking at 200 rev min−1 .

    Techniques: Expressing, Purification, Recombinant, SDS Page, Nucleic Acid Electrophoresis, Molecular Weight, Filtration

    3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated Gp78. (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by IPTG. Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).

    Journal: Molecular Biology of the Cell

    Article Title: p38 MAP kinase–dependent phosphorylation of the Gp78 E3 ubiquitin ligase controls ER–mitochondria association and mitochondria motility

    doi: 10.1091/mbc.E15-02-0120

    Figure Lengend Snippet: 3F3A mAb, but not anti-Gp78C, selectively recognizes dephosphorylated Gp78. (A) Cos7 cells were untransfected (Cos7 NT) or transfected with empty vector (PC3 EV), FLAG-tagged wild type (WT), S538A, phosphomimetic S538D or S538E, D536A mutant, or amino acid 533–541 deletion mutant (Δ3F3A) of Gp78. Cell lysates were immunoprecipitated with FLAG beads and probed with 3F3A, Gp78C, or FLAG mAbs. (B) Bacteria transformed with GST-tagged empty vector (GST-EV) or Gp78 C-terminal (GST-Gp78C) wild type (WT), S538A, S538D, S538E mutants, or amino acid 533–541 deletion mutant (Δ3F3A) were uninduced or induced by IPTG. Total bacterial lysates were dot blotted onto membranes and probed with 3F3A, Gp78C, or GST mAbs. (C) FLAG-tagged wild type of Gp78 (FLAG-Gp78-WT) or phosphorylation dominant-negative mutant (FLAG-Gp78 S538A) from Cos7 cells were immunoprecipitated and treated or not with CIAP and probed with 3F3A mAb, Gp78C, or FLAG mAb (±SEM; n = 3).

    Article Snippet: GST-Gp78C protein expression was induced by 1 mM isopropyl-β-d -thiogalactoside (IPTG) at 37°C for 4 h. pE-SUMO His-Gp78 was purchased from LifeSensors and the His-hGp78C induced with 0.4 mM IPTG at 16°C overnight and purified with His60 Ni Superflow Resin (Clontech). pcDNA3-Flag dominant-negative MKK6(K82A) and constitutively active MKK6 (Glu) were gifts from Roger Davis (University of Massachusetts, Worcester, MA; Addgene plasmids 13519 and 13518; ).

    Techniques: Transfection, Plasmid Preparation, Mutagenesis, Immunoprecipitation, Transformation Assay, Dominant Negative Mutation

    SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with IPTG for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl β-D-1-thiogalactopyranoside.

    Journal: Drug Design, Development and Therapy

    Article Title: Computational and nonglycosylated systems: a simpler approach for development of nanosized PEGylated proteins

    doi: 10.2147/DDDT.S98323

    Figure Lengend Snippet: SDS-Page and Western blot analysis. Notes: ( A ) The expression analyses of E31C and E89C ngEPO analogs, using 12% SDS-PAGE. Lane 1, SM0431 protein marker (Fermentas, Vilinius, Lithuania); lanes 2 and 3, samples from Escherichia coli after and before induction with IPTG for E31C analog; and lanes 4 and 5, samples from E. coli after and before induction with IPTG for E89C analog. ( B ) The Western blot analyses of expressed ngEPO analogs. Lane 1, SM0431 protein marker (Fermentas); lane 2, E. coli transformed with pET-26b vector; lane 3, E. coli transformed with E31C cDNA before induction with IPTG; lane 4, E. coli transformed with E31C cDNA after induction with IPTG; and lane 5, E. coli transformed with E89C cDNA after induction with IPTG. Abbreviations: MW, molecular weight; ngEPO, nonglycosylated erythropoietin; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IPTG, isopropyl β-D-1-thiogalactopyranoside.

    Article Snippet: The expression of analogs was induced using 1 mM isopropyl β-d -1-thiogalactopyranoside (IPTG) (Sigma-Aldrich, Munich, Germany) and analyzed using 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) using Mini-PROTEIN® Tetra Cell system (Bio-Rad Laboratories, Hercules, CA, USA); the gel was developed using the Coomassie blue staining technique.

    Techniques: SDS Page, Western Blot, Expressing, Marker, Transformation Assay, Positron Emission Tomography, Plasmid Preparation, Molecular Weight, Polyacrylamide Gel Electrophoresis

    SOD activities from S. aureus sod mutants. Lane 1, RN6390; lane 2, sodM mutant; lane 3, sodA mutant; lane 4, double ( sodM sodA ) mutant; lane 5, double mutant containing pCL15 sodM induced with IPTG; lane 6, double mutant containing pCL15 sodM uninduced.

    Journal: Journal of Bacteriology

    Article Title: Identification and Characterization of a Second Superoxide Dismutase Gene (sodM) from Staphylococcus aureus

    doi: 10.1128/JB.183.11.3399-3407.2001

    Figure Lengend Snippet: SOD activities from S. aureus sod mutants. Lane 1, RN6390; lane 2, sodM mutant; lane 3, sodA mutant; lane 4, double ( sodM sodA ) mutant; lane 5, double mutant containing pCL15 sodM induced with IPTG; lane 6, double mutant containing pCL15 sodM uninduced.

    Article Snippet: Expression of sodM was induced with IPTG (isopropyl-β- d -thiogalactopyranoside, 0.5 mM; Fisher Scientific, Fairlawn, N.J.) 1 h postinoculation, and cells were harvested 5 h later.

    Techniques: Mutagenesis

    Analysis of recombinant GST-muCCR5-NH2 terminal fusion protein. Mr: MW marker, 1, 2: products purified by glutathione sepharose, 3, 4: products not purified, 1, 3: expression products induced by IPTG

    Journal: World Journal of Gastroenterology

    Article Title: CCR5 gene expression in fulminant hepatitis and DTH in mice

    doi: 10.3748/wjg.v4.i1.14

    Figure Lengend Snippet: Analysis of recombinant GST-muCCR5-NH2 terminal fusion protein. Mr: MW marker, 1, 2: products purified by glutathione sepharose, 3, 4: products not purified, 1, 3: expression products induced by IPTG

    Article Snippet: The expression and purification of a GST fusion protein were induced with 0.1 M IPTG (isopropylthio-β-galactoside, Wako Pure Chemical Industries, LtD) for 5 h, put on glutathione-sepharose 4B affinity column (Pharmacia Biotech AB Upsala, Sweden), and then eluted with 5 mM of reduced glutathione.

    Techniques: Recombinant, Marker, Purification, Expressing

    Induction of p53 down regulates Bcl-3 protein levels in H1299 w/tp53 cells. (A) p53 represses Bcl-3 but not p100/p52 protein levels. Following 16 h of p53 induction by increasing concentrations of IPTG in H1299 w/tp53 cells, whole-cell lysates were prepared and the protein levels of p100/p52, Bcl-3, and β-actin control were analyzed by Western blot. (B) Bcl-3 down regulation is evident 12 h after IPTG induction of p53. p53 was induced in H1299 w/tp53 cells by 100 μM IPTG. Cells were harvested at the indicated times following IPTG treatment, and nuclear protein extracts were prepared and analyzed by Western blot for Bcl-3 levels. (C) Induction of p53 in H1299 w/tp53 cells does not repress Bcl-3 mRNA levels. RT-PCR analysis was performed by using two sets of primers specific to Bcl-3 (P1 and P2) or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: Induction of p53 down regulates Bcl-3 protein levels in H1299 w/tp53 cells. (A) p53 represses Bcl-3 but not p100/p52 protein levels. Following 16 h of p53 induction by increasing concentrations of IPTG in H1299 w/tp53 cells, whole-cell lysates were prepared and the protein levels of p100/p52, Bcl-3, and β-actin control were analyzed by Western blot. (B) Bcl-3 down regulation is evident 12 h after IPTG induction of p53. p53 was induced in H1299 w/tp53 cells by 100 μM IPTG. Cells were harvested at the indicated times following IPTG treatment, and nuclear protein extracts were prepared and analyzed by Western blot for Bcl-3 levels. (C) Induction of p53 in H1299 w/tp53 cells does not repress Bcl-3 mRNA levels. RT-PCR analysis was performed by using two sets of primers specific to Bcl-3 (P1 and P2) or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction

    p53-induced repression of cyclin D1 promoter activity can be prevented by Bcl-3 and the C terminus of p100. (A) Inhibition of cyclin D1 promoter activity by p53 can be reversed by expression of Bcl-3 or CT-p100. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of Bcl-3, CT-p100, or control RSV expression plasmid into H1299 w/tp53 cells. p53 expression was induced by 100 μM IPTG, and cells were harvested 16 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (B) Inhibition of cyclin D1 promoter activity by p53 is not prevented by p52. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of p52 or control RSV expression plasmid into H1299 w/tp53 cells. p53 expression was induced by 100 μM IPTG, and cells were harvested 16 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: p53-induced repression of cyclin D1 promoter activity can be prevented by Bcl-3 and the C terminus of p100. (A) Inhibition of cyclin D1 promoter activity by p53 can be reversed by expression of Bcl-3 or CT-p100. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of Bcl-3, CT-p100, or control RSV expression plasmid into H1299 w/tp53 cells. p53 expression was induced by 100 μM IPTG, and cells were harvested 16 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (B) Inhibition of cyclin D1 promoter activity by p53 is not prevented by p52. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of p52 or control RSV expression plasmid into H1299 w/tp53 cells. p53 expression was induced by 100 μM IPTG, and cells were harvested 16 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Activity Assay, Inhibition, Expressing, Luciferase, Plasmid Preparation, Activation Assay

    p53 selectively down regulates cyclin D1 expression. (A) Induction of p53 in H1299 w/tp53 cells represses cyclin D1 protein in a dose-dependent manner. Whole-cell lysates from H1299 w/tp53 cells, treated for 16 h with increasing concentrations of IPTG, were subjected to Western blot analysis by using antibodies to cyclins A, B1, D1, E, and a β-actin control as indicated. (B) Induction of p53 in H1299 w/tp53 cells represses cyclin D1 mRNA levels. RT-PCR analysis was performed by using primers specific to cyclin D1 or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: p53 selectively down regulates cyclin D1 expression. (A) Induction of p53 in H1299 w/tp53 cells represses cyclin D1 protein in a dose-dependent manner. Whole-cell lysates from H1299 w/tp53 cells, treated for 16 h with increasing concentrations of IPTG, were subjected to Western blot analysis by using antibodies to cyclins A, B1, D1, E, and a β-actin control as indicated. (B) Induction of p53 in H1299 w/tp53 cells represses cyclin D1 mRNA levels. RT-PCR analysis was performed by using primers specific to cyclin D1 or a GAPDH control, with total RNA prepared from H1299 w/tp53 cells treated for 16 h with IPTG.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

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

    p53 specifically inhibits cyclin D1 promoter activity. (A) Schematic diagram of the cyclin D1, cyclin D1 (−66), and cyclin D1 (−66 mut) luciferase reporter plasmids used. The relative positions of the previously described NF-κB binding sites are shown. luc, luciferase. (B) Relative luciferase activity of each of the cyclin D1-luciferase constructs. A quantity of 1.5 μg of each of the cyclin D1-luciferase reporter plasmids was transfected into H1299 w/tp53 cells. (C) Inhibition of cyclin D1 promoter activity by p53 requires the proximal κB site. A quantity of 1.5 μg of each of the cyclin D1-luciferase reporter plasmids was transfected into H1299 w/tp53 cells. p53 was induced by treatment with 100 μM IPTG, and cells were harvested 16 h after induction. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. (D) p21 does not inhibit cyclin D1 promoter activity in H1299 w/tp53 cells. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of RSV-p21 or control RSV plasmid. Results are expressed as relative luciferase activity. (E) p53 does not inhibit cyclin E promoter activity in H1299 w/tp53 cells. One and a half micrograms of cyclin E luciferase plasmid was transfected into H1299 w/tp53 cells, and p53 expression was induced by 100 μM IPTG. Cells were harvested 16 h after IPTG treatment, and results are expressed as relative luciferase activity.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: p53 specifically inhibits cyclin D1 promoter activity. (A) Schematic diagram of the cyclin D1, cyclin D1 (−66), and cyclin D1 (−66 mut) luciferase reporter plasmids used. The relative positions of the previously described NF-κB binding sites are shown. luc, luciferase. (B) Relative luciferase activity of each of the cyclin D1-luciferase constructs. A quantity of 1.5 μg of each of the cyclin D1-luciferase reporter plasmids was transfected into H1299 w/tp53 cells. (C) Inhibition of cyclin D1 promoter activity by p53 requires the proximal κB site. A quantity of 1.5 μg of each of the cyclin D1-luciferase reporter plasmids was transfected into H1299 w/tp53 cells. p53 was induced by treatment with 100 μM IPTG, and cells were harvested 16 h after induction. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. (D) p21 does not inhibit cyclin D1 promoter activity in H1299 w/tp53 cells. One and a half micrograms of cyclin D1-luciferase reporter plasmid was cotransfected with 1.5 μg of RSV-p21 or control RSV plasmid. Results are expressed as relative luciferase activity. (E) p53 does not inhibit cyclin E promoter activity in H1299 w/tp53 cells. One and a half micrograms of cyclin E luciferase plasmid was transfected into H1299 w/tp53 cells, and p53 expression was induced by 100 μM IPTG. Cells were harvested 16 h after IPTG treatment, and results are expressed as relative luciferase activity.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

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

    Down regulation of Bcl-3 and cyclin D1 by p53 in H1299 cells is dependent on phosphorylation of p53 at serine 15. (A) p53 expression in Saos-2 cells does not result in either the down regulation of Bcl-3 or cyclin D1. Saos-2 cells containing p53 under the control of the Tet-On-inducible system were harvested after 24 h of doxycycline treatment, and nuclear protein extracts were prepared. Western blot analysis of p53, p21, cyclin D1, and Bcl-3 was performed on p53-induced and control-treated samples. (B) p53 expressed in Saos-2 cells, in the absence of additional stimulation, is not phosphorylated at serine 15. Saos-2 cells containing p53 under the control of the Tet-On-inducible system were harvested at the indicated times following doxycycline treatment, and nuclear protein extracts were prepared. p53 and phospho-serine 15 p53 levels were then determined by Western blot analysis. (C) p53 is expressed to similar levels in both H1299 w/tp53 and H1299 S15A cells. Equivalent levels of protein from nuclear protein extracts prepared from H1299 w/tp53 and H1299 S15A cells treated with IPTG for 16 h were analyzed for p53 levels by Western blot. (D) Ser15Ala mutant p53 is unable to induce p21-luciferase activity or inhibit cyclin D1 promoter activity. One and a half micrograms of either p21-luciferase or cyclin D1-luciferase reporter plasmid was transfected into H1299/mutp53 cells. Cells were harvested following 16 h of IPTG treatment. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (E) Ser15Ala mutant p53 fails to down regulate Bcl-3 or cyclin D1 protein levels. Following 16 h of p53 induction by increasing concentrations of IPTG in H1299 S15A cells, whole-cell lysates were prepared, and the protein levels of Bcl-3 and cyclin D1 were analyzed by Western blot.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: Down regulation of Bcl-3 and cyclin D1 by p53 in H1299 cells is dependent on phosphorylation of p53 at serine 15. (A) p53 expression in Saos-2 cells does not result in either the down regulation of Bcl-3 or cyclin D1. Saos-2 cells containing p53 under the control of the Tet-On-inducible system were harvested after 24 h of doxycycline treatment, and nuclear protein extracts were prepared. Western blot analysis of p53, p21, cyclin D1, and Bcl-3 was performed on p53-induced and control-treated samples. (B) p53 expressed in Saos-2 cells, in the absence of additional stimulation, is not phosphorylated at serine 15. Saos-2 cells containing p53 under the control of the Tet-On-inducible system were harvested at the indicated times following doxycycline treatment, and nuclear protein extracts were prepared. p53 and phospho-serine 15 p53 levels were then determined by Western blot analysis. (C) p53 is expressed to similar levels in both H1299 w/tp53 and H1299 S15A cells. Equivalent levels of protein from nuclear protein extracts prepared from H1299 w/tp53 and H1299 S15A cells treated with IPTG for 16 h were analyzed for p53 levels by Western blot. (D) Ser15Ala mutant p53 is unable to induce p21-luciferase activity or inhibit cyclin D1 promoter activity. One and a half micrograms of either p21-luciferase or cyclin D1-luciferase reporter plasmid was transfected into H1299/mutp53 cells. Cells were harvested following 16 h of IPTG treatment. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (E) Ser15Ala mutant p53 fails to down regulate Bcl-3 or cyclin D1 protein levels. Following 16 h of p53 induction by increasing concentrations of IPTG in H1299 S15A cells, whole-cell lysates were prepared, and the protein levels of Bcl-3 and cyclin D1 were analyzed by Western blot.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Expressing, Western Blot, Mutagenesis, Luciferase, Activity Assay, Plasmid Preparation, Transfection, Activation Assay

    Induction of p53 expression in H1299 w/tp53 cells. (A) p53 induced in H1299 w/tp53 cells is phosphorylated at serine 15 and stimulates p21 expression. Western blot analysis of nuclear protein extracts showing the time course of induction of p53, p21, and phospho-serine 15-p53, following treatment with 100 μM IPTG in H1299 w/tp53 cells. (B) Induction of p53 in H1299 w/tp53 cells is dependent on the dose of IPTG. p53 and p21 expression was analyzed by Western blotting of nuclear protein extracts prepared from H1299 w/tp53 cells following 16 h of treatment with increasing concentrations of IPTG. (C) The level of p53 induced in H1299 w/tp53 is comparable to endogenous wild-type p53 levels in other cell lines. p53 expression was analyzed by Western blotting of nuclear protein extracts (15 μg) prepared from H1299 w/tp53 cells following 16 h of treatment and the other indicated cell lines. (D) Induction of p53 in H1299 w/tp53 cells results in a G 1 cell cycle arrest. Control cells or H1299 w/tp53 cells treated with 100 μM IPTG for 36 h were stained with propidium iodide and analyzed by using FACS analysis.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: Induction of p53 expression in H1299 w/tp53 cells. (A) p53 induced in H1299 w/tp53 cells is phosphorylated at serine 15 and stimulates p21 expression. Western blot analysis of nuclear protein extracts showing the time course of induction of p53, p21, and phospho-serine 15-p53, following treatment with 100 μM IPTG in H1299 w/tp53 cells. (B) Induction of p53 in H1299 w/tp53 cells is dependent on the dose of IPTG. p53 and p21 expression was analyzed by Western blotting of nuclear protein extracts prepared from H1299 w/tp53 cells following 16 h of treatment with increasing concentrations of IPTG. (C) The level of p53 induced in H1299 w/tp53 is comparable to endogenous wild-type p53 levels in other cell lines. p53 expression was analyzed by Western blotting of nuclear protein extracts (15 μg) prepared from H1299 w/tp53 cells following 16 h of treatment and the other indicated cell lines. (D) Induction of p53 in H1299 w/tp53 cells results in a G 1 cell cycle arrest. Control cells or H1299 w/tp53 cells treated with 100 μM IPTG for 36 h were stained with propidium iodide and analyzed by using FACS analysis.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Expressing, Western Blot, Staining, FACS

    Induction of p53 reduces p52/Bcl-3 complexes and increases p52 association with HDAC1. (A) Modulation of p52 complex formation by p53. Following 16 h of p53 induction by 100 μM IPTG in H1299 w/tp53 cells, nuclear protein extracts were prepared. Immunoprecipitation experiments were then performed with either a monoclonal or polyclonal antibody to p52, Bcl-3 antibody, or an immunoglobulin G control, from 100 μg of protein extract. The immunoprecipitated complex was then resolved by SDS-PAGE and immunoblotted with the indicated antibodies. Samples of input material are shown (10 μg). (B) Inhibition of HDAC activity abolishes p53-mediated repression of the cyclin D1 promoter. One and a half micrograms of each of the cyclin D1, cyclin D1 (−66), and cyclin D1 (−66 mut) luciferase reporter plasmids was transfected into H1299 w/tp53 cells. p53 was induced by treatment with 100 μM IPTG, and cells were harvested 16 h after induction. Cells were treated with 100 ng of TSA per ml for 16 h before harvesting as indicated. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (C) Inhibition of HDAC activity abolishes p53-mediated repression of endogenous cyclin D1. RT-PCR analysis was performed by using primers specific to cyclin D1 or a GAPDH control, with total RNA (100 ng) prepared from H1299 w/tp53 cells treated for 16 h with IPTG. Cells were treated with 100 ng of TSA per ml for 16 h before harvesting as indicated. (D) HDAC1 represses the cyclin D1 promoter in a κB element-dependent manner. The cyclin D1 (−66) or cyclin D1 (−66 mut) luciferase reporter plasmid (1.5 μg of each) was transfected into H1299 w/tp53 or U-2 OS cells together with 1 μg of HDAC1 expression plasmid. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: Induction of p53 reduces p52/Bcl-3 complexes and increases p52 association with HDAC1. (A) Modulation of p52 complex formation by p53. Following 16 h of p53 induction by 100 μM IPTG in H1299 w/tp53 cells, nuclear protein extracts were prepared. Immunoprecipitation experiments were then performed with either a monoclonal or polyclonal antibody to p52, Bcl-3 antibody, or an immunoglobulin G control, from 100 μg of protein extract. The immunoprecipitated complex was then resolved by SDS-PAGE and immunoblotted with the indicated antibodies. Samples of input material are shown (10 μg). (B) Inhibition of HDAC activity abolishes p53-mediated repression of the cyclin D1 promoter. One and a half micrograms of each of the cyclin D1, cyclin D1 (−66), and cyclin D1 (−66 mut) luciferase reporter plasmids was transfected into H1299 w/tp53 cells. p53 was induced by treatment with 100 μM IPTG, and cells were harvested 16 h after induction. Cells were treated with 100 ng of TSA per ml for 16 h before harvesting as indicated. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. luc, luciferase. (C) Inhibition of HDAC activity abolishes p53-mediated repression of endogenous cyclin D1. RT-PCR analysis was performed by using primers specific to cyclin D1 or a GAPDH control, with total RNA (100 ng) prepared from H1299 w/tp53 cells treated for 16 h with IPTG. Cells were treated with 100 ng of TSA per ml for 16 h before harvesting as indicated. (D) HDAC1 represses the cyclin D1 promoter in a κB element-dependent manner. The cyclin D1 (−66) or cyclin D1 (−66 mut) luciferase reporter plasmid (1.5 μg of each) was transfected into H1299 w/tp53 or U-2 OS cells together with 1 μg of HDAC1 expression plasmid. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Immunoprecipitation, SDS Page, Inhibition, Activity Assay, Luciferase, Transfection, Activation Assay, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Expressing

    Regulation of the cyclin D1 promoter by p52 and Bcl-3. (A) Schematic diagram of p100, p52, and the region of the C terminus of p100 used to construct the CT-p100 expression plasmid. (B) Effects of p52, Bcl-3, and CT-p100 expression on cyclin D1 promoter activity. One and a half micrograms of each of the indicated RSV expression plasmids was cotransfected with 1.5 μg of the indicated cyclin D1-luciferase reporter plasmids into H1299 w/tp53 cells in the absence of IPTG treatment. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. Western blot analysis of p52 expression in nuclear protein extracts from comparably transfected cells is shown inset. luc, luciferase. (C) CT-p100 does not inhibit TNF-α-induced NF-κB transcriptional activity. The 3× κB-luciferase reporter plasmid (1.5 μg) was cotransfected with 1.5 μg of either the CT-p100, SR-IκBα, or control RSV expression plasmid into H1299 w/tp53 cells in the absence of IPTG treatment. Luciferase activity was induced by treatment with 10 ng of TNF-α per ml, and cells were harvested 12 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Journal: Molecular and Cellular Biology

    Article Title: p53 Represses Cyclin D1 Transcription through Down Regulation of Bcl-3 and Inducing Increased Association of the p52 NF-?B Subunit with Histone Deacetylase 1

    doi: 10.1128/MCB.23.13.4713-4727.2003

    Figure Lengend Snippet: Regulation of the cyclin D1 promoter by p52 and Bcl-3. (A) Schematic diagram of p100, p52, and the region of the C terminus of p100 used to construct the CT-p100 expression plasmid. (B) Effects of p52, Bcl-3, and CT-p100 expression on cyclin D1 promoter activity. One and a half micrograms of each of the indicated RSV expression plasmids was cotransfected with 1.5 μg of the indicated cyclin D1-luciferase reporter plasmids into H1299 w/tp53 cells in the absence of IPTG treatment. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls. Western blot analysis of p52 expression in nuclear protein extracts from comparably transfected cells is shown inset. luc, luciferase. (C) CT-p100 does not inhibit TNF-α-induced NF-κB transcriptional activity. The 3× κB-luciferase reporter plasmid (1.5 μg) was cotransfected with 1.5 μg of either the CT-p100, SR-IκBα, or control RSV expression plasmid into H1299 w/tp53 cells in the absence of IPTG treatment. Luciferase activity was induced by treatment with 10 ng of TNF-α per ml, and cells were harvested 12 h later. Results are expressed as change in activation or repression ( n -fold) relative to levels seen in the relevant untreated cell controls.

    Article Snippet: Induction of p53 was achieved by treating cells with 20 to 100 μM IPTG (Melford Laboratories) for the indicated times.

    Techniques: Construct, Expressing, Plasmid Preparation, Activity Assay, Luciferase, Activation Assay, Western Blot, Transfection

    Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM IPTG on Mueller–Hinton broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).

    Journal: The EMBO Journal

    Article Title: Outer membrane composition of a lipopolysaccharide-deficient Neisseria meningitidis mutant

    doi: 10.1093/emboj/20.24.6937

    Figure Lengend Snippet: Fig. 7. Silver-stained Tricine–SDS–PAGE gel showing the LPS pattern of proteinase K-treated whole-cell lysates of HA3003 grown in the absence (a) or presence (b) of 1 mM IPTG on Mueller–Hinton broth (MHB), Gonococcal Agar (GC) or Meningococcal Medium (Men Med).

    Article Snippet: To induce the tac promoter, strains were grown in Mueller–Hinton broth or Meningococcal Medium in the presence of 1 mM IPTG (Roche).

    Techniques: Staining, SDS Page