n dna ligase Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs n dna ligase
    Determinants of Thermococcus Okazaki fragment maturation. Panel I , a schematic of the Okazaki fragment maturation experiment is shown. The processing substrate was prepared by annealing a 5′-TAM-labeled extension primer (shaded black ) and 3′-FAM-labeled blocking oligonucleotide (shaded blue ) to ssM13 as described in the text. <t>9°N</t> polB, polD, PCNA/RFC, Fen1, and <t>DNA</t> ligase were incubated with the processing substrate at 60 °C for 15 min. Processed products (103 nt) result from complete Okazaki fragment maturation. B , Okazaki fragment maturation assays were performed without any proteins added, all proteins added (9°N polB, polD, PCNA/RFC, Fen1, and DNA ligase), or in reactions that omit one protein as noted in the figure. Reaction products were analyzed by capillary electrophoresis. 5′-TAM-labeled extension primer is shaded black and 3′-FAM-labeled blocking oligonucleotide is shaded blue . Processed products (103 nt) result from a complete Okazaki fragment maturation assay and are labeled with both 5′-TAM ( black ) and 3′-FAM ( blue ). C, processed products from reactions omitting various replication proteins in panel B were quantitated and plotted. The data shown are averages with standard deviations from three independent experiments.
    N Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n dna ligase/product/New England Biolabs
    Average 99 stars, based on 68 article reviews
    Price from $9.99 to $1999.99
    n dna ligase - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Millipore dna fragments
    Crystal structure of the human <t>CENP-A/H3.3</t> nucleosome. (a) The CENP-A/H3.3 nucleosome structure is presented. CENP-A and H3.3 molecules are colored red and blue, respectively. The 2mFo - DFc maps of the two <t>DNA</t> end regions of the CENP-A/H3.3 were calculated and contoured at the 2.0σ level. (b) Close-up views of the CENP-A αN helix, the H3.3 αN helix, the CENP-A Ser68 residue, the H3.3 Gln68 residue, the CENP-A His104 residue, and the H3.3 Gly102 residue. Electron density maps are presented at the 1.5σ level.
    Dna Fragments, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6627 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna fragments/product/Millipore
    Average 99 stars, based on 6627 article reviews
    Price from $9.99 to $1999.99
    dna fragments - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Millipore dna segment
    The C-terminal tryptophan residue (W180) of <t>FAAP20</t> outside the compact UBZ module plays an indispensable role in ubiquitin recognition by FAAP20 in vitro and in efficient ICL <t>DNA</t> repair in vivo . ( A ) Measurement of binding affinity between WT FAAP20 and WT ubiquitin by ITC. ( B ) Mutation of W180A in FAAP20 abolished ubiquitin binding in ITC measurements. ( C ) U2OS cells stably expressing siRNA-resistant FAAP20 wild-type, C147A and C150A (CA), or W180A (WA) were transfected with siRNA against FAAP20 (siF20) for 48 h, and cell viability was determined 6 days after the treatment of indicated doses of mitomycin C. ( D ) Immunoblot analysis of U2OS cells in (C) harvested 72 h after siRNA treatment.
    Dna Segment, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1563 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna segment/product/Millipore
    Average 99 stars, based on 1563 article reviews
    Price from $9.99 to $1999.99
    dna segment - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs 9°n dna ligase
    The C-terminal tryptophan residue (W180) of <t>FAAP20</t> outside the compact UBZ module plays an indispensable role in ubiquitin recognition by FAAP20 in vitro and in efficient ICL <t>DNA</t> repair in vivo . ( A ) Measurement of binding affinity between WT FAAP20 and WT ubiquitin by ITC. ( B ) Mutation of W180A in FAAP20 abolished ubiquitin binding in ITC measurements. ( C ) U2OS cells stably expressing siRNA-resistant FAAP20 wild-type, C147A and C150A (CA), or W180A (WA) were transfected with siRNA against FAAP20 (siF20) for 48 h, and cell viability was determined 6 days after the treatment of indicated doses of mitomycin C. ( D ) Immunoblot analysis of U2OS cells in (C) harvested 72 h after siRNA treatment.
    9°N Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/9°n dna ligase/product/New England Biolabs
    Average 99 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    9°n dna ligase - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Determinants of Thermococcus Okazaki fragment maturation. Panel I , a schematic of the Okazaki fragment maturation experiment is shown. The processing substrate was prepared by annealing a 5′-TAM-labeled extension primer (shaded black ) and 3′-FAM-labeled blocking oligonucleotide (shaded blue ) to ssM13 as described in the text. 9°N polB, polD, PCNA/RFC, Fen1, and DNA ligase were incubated with the processing substrate at 60 °C for 15 min. Processed products (103 nt) result from complete Okazaki fragment maturation. B , Okazaki fragment maturation assays were performed without any proteins added, all proteins added (9°N polB, polD, PCNA/RFC, Fen1, and DNA ligase), or in reactions that omit one protein as noted in the figure. Reaction products were analyzed by capillary electrophoresis. 5′-TAM-labeled extension primer is shaded black and 3′-FAM-labeled blocking oligonucleotide is shaded blue . Processed products (103 nt) result from a complete Okazaki fragment maturation assay and are labeled with both 5′-TAM ( black ) and 3′-FAM ( blue ). C, processed products from reactions omitting various replication proteins in panel B were quantitated and plotted. The data shown are averages with standard deviations from three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: The Roles of Family B and D DNA Polymerases in Thermococcus Species 9°N Okazaki Fragment Maturation *

    doi: 10.1074/jbc.M115.638130

    Figure Lengend Snippet: Determinants of Thermococcus Okazaki fragment maturation. Panel I , a schematic of the Okazaki fragment maturation experiment is shown. The processing substrate was prepared by annealing a 5′-TAM-labeled extension primer (shaded black ) and 3′-FAM-labeled blocking oligonucleotide (shaded blue ) to ssM13 as described in the text. 9°N polB, polD, PCNA/RFC, Fen1, and DNA ligase were incubated with the processing substrate at 60 °C for 15 min. Processed products (103 nt) result from complete Okazaki fragment maturation. B , Okazaki fragment maturation assays were performed without any proteins added, all proteins added (9°N polB, polD, PCNA/RFC, Fen1, and DNA ligase), or in reactions that omit one protein as noted in the figure. Reaction products were analyzed by capillary electrophoresis. 5′-TAM-labeled extension primer is shaded black and 3′-FAM-labeled blocking oligonucleotide is shaded blue . Processed products (103 nt) result from a complete Okazaki fragment maturation assay and are labeled with both 5′-TAM ( black ) and 3′-FAM ( blue ). C, processed products from reactions omitting various replication proteins in panel B were quantitated and plotted. The data shown are averages with standard deviations from three independent experiments.

    Article Snippet: Enzymes and Reagents All restriction endonucleases, modifying enzymes, polB (9°Nm DNA polymerase; 9°N/E143D), 9°N DNA ligase, T4 DNA ligase, nucleotides, and single-stranded M13mp18 DNA (ssM13) were from New England Biolabs, Inc. (Ipswich, MA).

    Techniques: Labeling, Blocking Assay, Incubation, Electrophoresis

    Simplified models of Okazaki fragment maturation in bacteria ( A ), Eukarya ( B ), and Thermococcus species 9°N ( C ). A, in bacteria, pol III synthesizes the lagging strand. pol I replaces pol III to complete Okazaki fragment maturation. pol I 5′-3′ exonuclease removes the RNA primer as its DNA polymerase activity fills the gap. DNA ligase seals the Okazaki fragments. B, the eukaryal lagging strand DNA polymerase, pol δ, strand displacement activity generates a flap for Fen1 cleavage. DNA ligase seals the Okazaki fragments. C, polD synthesizes the lagging strand and stops at a downstream Okazaki fragment. polB replaces polD and its strand displacement activity generates a flap for Fen1 cleavage. DNA ligase seals the Okazaki fragments. Further details are described in the text.

    Journal: The Journal of Biological Chemistry

    Article Title: The Roles of Family B and D DNA Polymerases in Thermococcus Species 9°N Okazaki Fragment Maturation *

    doi: 10.1074/jbc.M115.638130

    Figure Lengend Snippet: Simplified models of Okazaki fragment maturation in bacteria ( A ), Eukarya ( B ), and Thermococcus species 9°N ( C ). A, in bacteria, pol III synthesizes the lagging strand. pol I replaces pol III to complete Okazaki fragment maturation. pol I 5′-3′ exonuclease removes the RNA primer as its DNA polymerase activity fills the gap. DNA ligase seals the Okazaki fragments. B, the eukaryal lagging strand DNA polymerase, pol δ, strand displacement activity generates a flap for Fen1 cleavage. DNA ligase seals the Okazaki fragments. C, polD synthesizes the lagging strand and stops at a downstream Okazaki fragment. polB replaces polD and its strand displacement activity generates a flap for Fen1 cleavage. DNA ligase seals the Okazaki fragments. Further details are described in the text.

    Article Snippet: Enzymes and Reagents All restriction endonucleases, modifying enzymes, polB (9°Nm DNA polymerase; 9°N/E143D), 9°N DNA ligase, T4 DNA ligase, nucleotides, and single-stranded M13mp18 DNA (ssM13) were from New England Biolabs, Inc. (Ipswich, MA).

    Techniques: Activity Assay

    Crystal structure of the human CENP-A/H3.3 nucleosome. (a) The CENP-A/H3.3 nucleosome structure is presented. CENP-A and H3.3 molecules are colored red and blue, respectively. The 2mFo - DFc maps of the two DNA end regions of the CENP-A/H3.3 were calculated and contoured at the 2.0σ level. (b) Close-up views of the CENP-A αN helix, the H3.3 αN helix, the CENP-A Ser68 residue, the H3.3 Gln68 residue, the CENP-A His104 residue, and the H3.3 Gly102 residue. Electron density maps are presented at the 1.5σ level.

    Journal: Scientific Reports

    Article Title: Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3

    doi: 10.1038/srep07115

    Figure Lengend Snippet: Crystal structure of the human CENP-A/H3.3 nucleosome. (a) The CENP-A/H3.3 nucleosome structure is presented. CENP-A and H3.3 molecules are colored red and blue, respectively. The 2mFo - DFc maps of the two DNA end regions of the CENP-A/H3.3 were calculated and contoured at the 2.0σ level. (b) Close-up views of the CENP-A αN helix, the H3.3 αN helix, the CENP-A Ser68 residue, the H3.3 Gln68 residue, the CENP-A His104 residue, and the H3.3 Gly102 residue. Electron density maps are presented at the 1.5σ level.

    Article Snippet: Purification of the human CENP-C fragment The DNA fragment encoding human CENP-C(426–537) was ligated between the NdeI and BamHI sites of the pET15b (Novagen) expression vector.

    Techniques:

    The DNA end close to CENP-A is asymmetrically flexible in the CENP-A/H3.3 nucleosome. (a) MNase assay. The H3.3, CENP-A, and CENP-A/H3.3 nucleosomes were treated with MNase (0, 0.3, 0.5 and 0.7 units), and the resulting DNA fragments were analyzed by native PAGE. The gel image shown is a representative of four independent experiments, in which similar results were obtained. (b) ExoIII assay. The H3.3, CENP-A, or CENP-A/H3.3 nucleosomes were incubated with or without 2.0 units of ExoIII for 2.5, 5 and 7.5 minutes at 37°C, and the resultant DNA fragments were extracted and analyzed by 14% denaturing PAGE with 7 M urea. The gel image shown is a representative of three independent experiments, in which similar results were obtained.

    Journal: Scientific Reports

    Article Title: Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3

    doi: 10.1038/srep07115

    Figure Lengend Snippet: The DNA end close to CENP-A is asymmetrically flexible in the CENP-A/H3.3 nucleosome. (a) MNase assay. The H3.3, CENP-A, and CENP-A/H3.3 nucleosomes were treated with MNase (0, 0.3, 0.5 and 0.7 units), and the resulting DNA fragments were analyzed by native PAGE. The gel image shown is a representative of four independent experiments, in which similar results were obtained. (b) ExoIII assay. The H3.3, CENP-A, or CENP-A/H3.3 nucleosomes were incubated with or without 2.0 units of ExoIII for 2.5, 5 and 7.5 minutes at 37°C, and the resultant DNA fragments were extracted and analyzed by 14% denaturing PAGE with 7 M urea. The gel image shown is a representative of three independent experiments, in which similar results were obtained.

    Article Snippet: Purification of the human CENP-C fragment The DNA fragment encoding human CENP-C(426–537) was ligated between the NdeI and BamHI sites of the pET15b (Novagen) expression vector.

    Techniques: Clear Native PAGE, Incubation, Polyacrylamide Gel Electrophoresis

    Stability of the CENP-A/H3.3 nucleosome. (a) Schematic representation of the thermal stability assay. (b) Thermal stability curve of the H3.3 nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the H3.3 stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (c) A thermal stability curve of the CENP-A nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the CENP-A stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (d) Tetrasomes, reconstituted with the H3.3-H4 tetramer or the CENP-A-H4 tetramer and DNA, were analyzed by 6% native PAGE. Lane 1 indicates the H3.3 tetrasome before incubation. Lanes 2, 3, 4, 5, and 6 indicate the H3.3 tetrasomes after 25°C, 35°C, 45°C, 55°C, and 65°C incubations, respectively. Lane 7 indicates the CENP-A tetrasome before incubation. Lanes 8, 9, 10, 11, and 12 indicate the CENP-A tetrasomes after 25°C, 35°C, 45°C, 55°C, and 65°C incubations, respectively. DNA was visualized by ethidium bromide staining. The gel image is a representative of seven independent experiments with similar results. (e) A thermal stability curve of the CENP-A/H3.3 nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the CENP-A/H3.3 stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (f) The H3.3, CENP-A, and CENP-A/H3.3 nucleosomes were incubated for 1 min at each temperature from 25°C, and the samples at 65°C, 72°C, 79°C, and 86°C were analyzed by 6% native PAGE. DNA was visualized by ethidium bromide staining. The gel image is a representative of three independent experiments with similar results.

    Journal: Scientific Reports

    Article Title: Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3

    doi: 10.1038/srep07115

    Figure Lengend Snippet: Stability of the CENP-A/H3.3 nucleosome. (a) Schematic representation of the thermal stability assay. (b) Thermal stability curve of the H3.3 nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the H3.3 stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (c) A thermal stability curve of the CENP-A nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the CENP-A stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (d) Tetrasomes, reconstituted with the H3.3-H4 tetramer or the CENP-A-H4 tetramer and DNA, were analyzed by 6% native PAGE. Lane 1 indicates the H3.3 tetrasome before incubation. Lanes 2, 3, 4, 5, and 6 indicate the H3.3 tetrasomes after 25°C, 35°C, 45°C, 55°C, and 65°C incubations, respectively. Lane 7 indicates the CENP-A tetrasome before incubation. Lanes 8, 9, 10, 11, and 12 indicate the CENP-A tetrasomes after 25°C, 35°C, 45°C, 55°C, and 65°C incubations, respectively. DNA was visualized by ethidium bromide staining. The gel image is a representative of seven independent experiments with similar results. (e) A thermal stability curve of the CENP-A/H3.3 nucleosome. The fluorescence intensity was plotted against each temperature (from 55°C to 90°C). The derivative values of the CENP-A/H3.3 stability curve presented in the upper panel are plotted against the temperatures (bottom panel). Means ± s.d. ( n = 3) are shown. (f) The H3.3, CENP-A, and CENP-A/H3.3 nucleosomes were incubated for 1 min at each temperature from 25°C, and the samples at 65°C, 72°C, 79°C, and 86°C were analyzed by 6% native PAGE. DNA was visualized by ethidium bromide staining. The gel image is a representative of three independent experiments with similar results.

    Article Snippet: Purification of the human CENP-C fragment The DNA fragment encoding human CENP-C(426–537) was ligated between the NdeI and BamHI sites of the pET15b (Novagen) expression vector.

    Techniques: Stability Assay, Fluorescence, Clear Native PAGE, Incubation, Staining

    The C-terminal tryptophan residue (W180) of FAAP20 outside the compact UBZ module plays an indispensable role in ubiquitin recognition by FAAP20 in vitro and in efficient ICL DNA repair in vivo . ( A ) Measurement of binding affinity between WT FAAP20 and WT ubiquitin by ITC. ( B ) Mutation of W180A in FAAP20 abolished ubiquitin binding in ITC measurements. ( C ) U2OS cells stably expressing siRNA-resistant FAAP20 wild-type, C147A and C150A (CA), or W180A (WA) were transfected with siRNA against FAAP20 (siF20) for 48 h, and cell viability was determined 6 days after the treatment of indicated doses of mitomycin C. ( D ) Immunoblot analysis of U2OS cells in (C) harvested 72 h after siRNA treatment.

    Journal: Nucleic Acids Research

    Article Title: Ubiquitin recognition by FAAP20 expands the complex interface beyond the canonical UBZ domain

    doi: 10.1093/nar/gku1153

    Figure Lengend Snippet: The C-terminal tryptophan residue (W180) of FAAP20 outside the compact UBZ module plays an indispensable role in ubiquitin recognition by FAAP20 in vitro and in efficient ICL DNA repair in vivo . ( A ) Measurement of binding affinity between WT FAAP20 and WT ubiquitin by ITC. ( B ) Mutation of W180A in FAAP20 abolished ubiquitin binding in ITC measurements. ( C ) U2OS cells stably expressing siRNA-resistant FAAP20 wild-type, C147A and C150A (CA), or W180A (WA) were transfected with siRNA against FAAP20 (siF20) for 48 h, and cell viability was determined 6 days after the treatment of indicated doses of mitomycin C. ( D ) Immunoblot analysis of U2OS cells in (C) harvested 72 h after siRNA treatment.

    Article Snippet: Protein constructs and cloning The DNA sequence of residues 140–180 of human FAAP20 isoform 2 was synthesized; the polymerase chain reaction amplified DNA was double digested and ligated into a modified pET15b vector (EMD Biosciences, Inc.) between the NdeI and XhoI restriction sites.

    Techniques: In Vitro, In Vivo, Binding Assay, Mutagenesis, Stable Transfection, Expressing, Transfection

    Protein composition and specificity of DNA binding activity at the Rgs4 AP-1R sequence in vitro . Representative chemiluminescent images of EMSAs showing bands of biotin-labeled oligonucleotide probe shifted in the presence of nuclear protein extracts (1.2 μg) from either pineal gland ( P ) or brain cortex ( Cx ). A , note that two shifted bands ( upper and lower arrows ) are observed in the pineal gland, whereas only the equivalent upper band is observed in the cortex together with two additional slower migrating bands ( arrowheads ). Note that both EMSA bands are more abundant in pineal samples extracted at 12.00 versus 24.00 h. Unbound (free) probe is indicated at the gel base. B , summated results of multiple EMSAs comparing the abundance of the upper and lower shifted bands in pineal glands sampled at either 12.00 and 24.00 h. Values are fold-difference compared with the level at 12.00 h (mean ± S.E., n = 6, *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Selective Genomic Targeting by FRA-2/FOSL2 Transcription Factor

    doi: 10.1074/jbc.M110.201996

    Figure Lengend Snippet: Protein composition and specificity of DNA binding activity at the Rgs4 AP-1R sequence in vitro . Representative chemiluminescent images of EMSAs showing bands of biotin-labeled oligonucleotide probe shifted in the presence of nuclear protein extracts (1.2 μg) from either pineal gland ( P ) or brain cortex ( Cx ). A , note that two shifted bands ( upper and lower arrows ) are observed in the pineal gland, whereas only the equivalent upper band is observed in the cortex together with two additional slower migrating bands ( arrowheads ). Note that both EMSA bands are more abundant in pineal samples extracted at 12.00 versus 24.00 h. Unbound (free) probe is indicated at the gel base. B , summated results of multiple EMSAs comparing the abundance of the upper and lower shifted bands in pineal glands sampled at either 12.00 and 24.00 h. Values are fold-difference compared with the level at 12.00 h (mean ± S.E., n = 6, *, p

    Article Snippet: These fragments were amplified (Extract N′Amp, Sigma) from rat genomic DNA using PCR primers (Rgs4 -167F, Rgs4 -R, Rgs4 -m167F, Rgs4 -426F, and Rgs4 -m426F; supplemental Table S1 ).

    Techniques: Binding Assay, Activity Assay, Sequencing, In Vitro, Labeling

    Association of FRA-2 with Rgs4 promoter sequence in vivo . ChIP assays were conducted using chromatin extracted from rat pineal glands; gene promoter sequences were amplified by PCR and visualized by ethidium bromide staining of agarose gels. A , ChIP analysis reveals enrichment of pineal chromatin when precipitated with FRA-2 and RNA polymerase II ( Pol II ) antisera compared with a FRA-2 preimmune serum and IgG. IN , input chromatin; Con , water PCR control. Note that input chromatin is diluted relative to ChIP chromatin. A parallel assay conducted with primers specific for the control β-actin gene revealed enrichment only with polymerase II antisera. B , summated data from multiple Rgs4 ChIP assay. Values are expressed as % of input DNA (mean ± S.E. n = 3 assays from three individual groups of rats). Group means that are significantly different ( p

    Journal: The Journal of Biological Chemistry

    Article Title: Selective Genomic Targeting by FRA-2/FOSL2 Transcription Factor

    doi: 10.1074/jbc.M110.201996

    Figure Lengend Snippet: Association of FRA-2 with Rgs4 promoter sequence in vivo . ChIP assays were conducted using chromatin extracted from rat pineal glands; gene promoter sequences were amplified by PCR and visualized by ethidium bromide staining of agarose gels. A , ChIP analysis reveals enrichment of pineal chromatin when precipitated with FRA-2 and RNA polymerase II ( Pol II ) antisera compared with a FRA-2 preimmune serum and IgG. IN , input chromatin; Con , water PCR control. Note that input chromatin is diluted relative to ChIP chromatin. A parallel assay conducted with primers specific for the control β-actin gene revealed enrichment only with polymerase II antisera. B , summated data from multiple Rgs4 ChIP assay. Values are expressed as % of input DNA (mean ± S.E. n = 3 assays from three individual groups of rats). Group means that are significantly different ( p

    Article Snippet: These fragments were amplified (Extract N′Amp, Sigma) from rat genomic DNA using PCR primers (Rgs4 -167F, Rgs4 -R, Rgs4 -m167F, Rgs4 -426F, and Rgs4 -m426F; supplemental Table S1 ).

    Techniques: Sequencing, In Vivo, Chromatin Immunoprecipitation, Amplification, Polymerase Chain Reaction, Staining

    Mechanistic basis for nocturnal Rgs4 down-regulation by FRA-2. A , CBP is associated with the Rgs4 promoter. Chemiluminescent image of a representative EMSAs showing bands of biotin-labeled oligonucleotide probe shifted in the presence of nuclear protein extracts (1.2 μg) from pineal gland ( P ). The shifted bands are partially abrogated in the presence of a CREB-1 antisera (sc-186) and completely abrogated in the presence of CBP antisera (A22). A rabbit IgG was used in the Control lane. B , ChIP analysis revealed increased nocturnal occupancy of CBP at the Rgs4 promoter in DN-FRA-2 transgenic ( TG ) rat pineal glands sampled at night (24.00 h) compared with wild-type ( WT ) controls. Data were quantified by QPCR, expressed as % of input DNA (mean ± S.E.) and summated in histograms ( n = 4 assays from four individual groups of rats, p

    Journal: The Journal of Biological Chemistry

    Article Title: Selective Genomic Targeting by FRA-2/FOSL2 Transcription Factor

    doi: 10.1074/jbc.M110.201996

    Figure Lengend Snippet: Mechanistic basis for nocturnal Rgs4 down-regulation by FRA-2. A , CBP is associated with the Rgs4 promoter. Chemiluminescent image of a representative EMSAs showing bands of biotin-labeled oligonucleotide probe shifted in the presence of nuclear protein extracts (1.2 μg) from pineal gland ( P ). The shifted bands are partially abrogated in the presence of a CREB-1 antisera (sc-186) and completely abrogated in the presence of CBP antisera (A22). A rabbit IgG was used in the Control lane. B , ChIP analysis revealed increased nocturnal occupancy of CBP at the Rgs4 promoter in DN-FRA-2 transgenic ( TG ) rat pineal glands sampled at night (24.00 h) compared with wild-type ( WT ) controls. Data were quantified by QPCR, expressed as % of input DNA (mean ± S.E.) and summated in histograms ( n = 4 assays from four individual groups of rats, p

    Article Snippet: These fragments were amplified (Extract N′Amp, Sigma) from rat genomic DNA using PCR primers (Rgs4 -167F, Rgs4 -R, Rgs4 -m167F, Rgs4 -426F, and Rgs4 -m426F; supplemental Table S1 ).

    Techniques: Labeling, Chromatin Immunoprecipitation, Transgenic Assay, Real-time Polymerase Chain Reaction