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  • 99
    Thermo Fisher fugene hd transfection reagent
    Fugene Hd Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene hd transfection reagent/product/Thermo Fisher
    Average 99 stars, based on 150 article reviews
    Price from $9.99 to $1999.99
    fugene hd transfection reagent - by Bioz Stars, 2020-09
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    99
    Promega fugene hd
    Comparison of major commercially available transfection reagents using described flow cytometric method that quantifies transfection efficiency. 293T cells underwent chemical transfection using labeled or standard transfected nucleic acids (pNL4-3 or mCherry plasmids) and different commercially available transfection reagents as described in Methods. Gating on viable cells was performed as shown in Fig 1 . Each experiment was performed in triplicates and six independent times (6 per plasmid and 12 in total) with specific (un)labelled nucleic acids. Data in each experiment were normalized by the average of the experimental control (standard transfected nucleic acids) in each experiment and were then pooled together. This approach increases statistical power while taking into consideration the inherent differences in transfection efficiency among different transfection methods (different chemical transfection reagents and plasmids). The non-parametric statistical Mann-Whitney test was used for comparisons between groups. Median and interquartile range (IQR) are shown. The transfection efficiency was assessed by both amount of labelled DNA (A) and protein (B). The use of TransIT-X2 and Jet Prime gave the best (and comparable) transfection efficiencies, whereas the efficiency was reduced with <t>Lipofectamine</t> 2000 and <t>Fugene</t> HD. There was a major decrease in the amount of detectable labelled DNA with Lipofectamine 2000 and Fugene HD.
    Fugene Hd, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 16287 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene hd/product/Promega
    Average 99 stars, based on 16287 article reviews
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    99
    Promega fugene
    Rb phosphorylation at S807 is required for association with Bax. Rb mutant plasmids were generated as described in the Materials and Methods section. Rb-negative <t>C33A</t> cells were transfected using <t>Fugene</t> (Promega). ( A ) Rb plasmids expressing either S807A or S811A alanine mutants were transfected into cells and 48 hours later cell lysates were utilized in GST-Bax fusion protein pull down assays. Proteins associated with GST-Bax were analyzed by immunoblotting with Rb antibodies. ( B ) Rb plasmids expressing either S807A or S807E mutants were transfected into C33A cells and 48 hours later co-immunoprecipitation with Bax antibodies was performed. Immunoprecipitates were analyzed by immunoblotting with Rb and Bax antibodies. Equivalent expression of the Rb mutant proteins in the C33A cells is verified by immunoblotting (lower panel). Data shown is representative of three independent experiments.
    Fugene, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 4807 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene/product/Promega
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    fugene - by Bioz Stars, 2020-09
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    92
    Boehringer Mannheim fugene 6 transfection reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene 6 Transfection Reagent, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 558 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6 transfection reagent/product/Boehringer Mannheim
    Average 92 stars, based on 558 article reviews
    Price from $9.99 to $1999.99
    fugene 6 transfection reagent - by Bioz Stars, 2020-09
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    92
    Thermo Fisher fugene transfection reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene transfection reagent/product/Thermo Fisher
    Average 92 stars, based on 95 article reviews
    Price from $9.99 to $1999.99
    fugene transfection reagent - by Bioz Stars, 2020-09
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    Image Search Results


    Comparison of major commercially available transfection reagents using described flow cytometric method that quantifies transfection efficiency. 293T cells underwent chemical transfection using labeled or standard transfected nucleic acids (pNL4-3 or mCherry plasmids) and different commercially available transfection reagents as described in Methods. Gating on viable cells was performed as shown in Fig 1 . Each experiment was performed in triplicates and six independent times (6 per plasmid and 12 in total) with specific (un)labelled nucleic acids. Data in each experiment were normalized by the average of the experimental control (standard transfected nucleic acids) in each experiment and were then pooled together. This approach increases statistical power while taking into consideration the inherent differences in transfection efficiency among different transfection methods (different chemical transfection reagents and plasmids). The non-parametric statistical Mann-Whitney test was used for comparisons between groups. Median and interquartile range (IQR) are shown. The transfection efficiency was assessed by both amount of labelled DNA (A) and protein (B). The use of TransIT-X2 and Jet Prime gave the best (and comparable) transfection efficiencies, whereas the efficiency was reduced with Lipofectamine 2000 and Fugene HD. There was a major decrease in the amount of detectable labelled DNA with Lipofectamine 2000 and Fugene HD.

    Journal: PLoS ONE

    Article Title: A novel rapid and reproducible flow cytometric method for optimization of transfection efficiency in cells

    doi: 10.1371/journal.pone.0182941

    Figure Lengend Snippet: Comparison of major commercially available transfection reagents using described flow cytometric method that quantifies transfection efficiency. 293T cells underwent chemical transfection using labeled or standard transfected nucleic acids (pNL4-3 or mCherry plasmids) and different commercially available transfection reagents as described in Methods. Gating on viable cells was performed as shown in Fig 1 . Each experiment was performed in triplicates and six independent times (6 per plasmid and 12 in total) with specific (un)labelled nucleic acids. Data in each experiment were normalized by the average of the experimental control (standard transfected nucleic acids) in each experiment and were then pooled together. This approach increases statistical power while taking into consideration the inherent differences in transfection efficiency among different transfection methods (different chemical transfection reagents and plasmids). The non-parametric statistical Mann-Whitney test was used for comparisons between groups. Median and interquartile range (IQR) are shown. The transfection efficiency was assessed by both amount of labelled DNA (A) and protein (B). The use of TransIT-X2 and Jet Prime gave the best (and comparable) transfection efficiencies, whereas the efficiency was reduced with Lipofectamine 2000 and Fugene HD. There was a major decrease in the amount of detectable labelled DNA with Lipofectamine 2000 and Fugene HD.

    Article Snippet: Transfection reagents The following commercially available transfection reagents were purchased: TransIT-X2 (Mirus, Madison, WI), Jet Prime (Polyplus, France), Lipofectamine 2000 (Thermo Fisher, Waltham, MA) and Fugene HD (Promega, Madison, WI).

    Techniques: Transfection, Flow Cytometry, Cytometry, Labeling, Plasmid Preparation, MANN-WHITNEY

    Efficiency of DAI knockdown in primary murine microglia and astrocytes by small-interfering RNA . Microglia (Mg) and astrocytes (Ast) were untreated (Con) or transfected with a combination of three different siRNA duplexes targeting murine DAI using FuGENE HD transfection reagent. At 72 hours post-transfection, untreated and transfected cells were exposed to HSV-1 (MOI of 10) for 24 hours and DAI knockdown was confirmed in whole cell lysates by immunoblot analysis. Expression of a non-specific protein band observed following Coomassie blue staining is shown as a loading control (lc). For comparison purposes, DAI expression in murine small intestine tissue is shown (+).

    Journal: Journal of Neuroinflammation

    Article Title: A role for DNA-dependent activator of interferon regulatory factor in the recognition of herpes simplex virus type 1 by glial cells

    doi: 10.1186/1742-2094-8-99

    Figure Lengend Snippet: Efficiency of DAI knockdown in primary murine microglia and astrocytes by small-interfering RNA . Microglia (Mg) and astrocytes (Ast) were untreated (Con) or transfected with a combination of three different siRNA duplexes targeting murine DAI using FuGENE HD transfection reagent. At 72 hours post-transfection, untreated and transfected cells were exposed to HSV-1 (MOI of 10) for 24 hours and DAI knockdown was confirmed in whole cell lysates by immunoblot analysis. Expression of a non-specific protein band observed following Coomassie blue staining is shown as a loading control (lc). For comparison purposes, DAI expression in murine small intestine tissue is shown (+).

    Article Snippet: In vitro stimulation of microglia and astrocytes with the DAI ligand, B-DNA Poly(dA:dT) double-stranded B-DNA (InvivoGen, San Diego, CA) was directly introduced into microglia and astrocytes at concentrations of 3 μg/ml and 6 μg/ml using FuGENE HD Transfection Reagent (Promega, Madison, WI) according to the manufacturer's instructions.

    Techniques: Small Interfering RNA, AST Assay, Transfection, Expressing, Staining

    The role of miR-208a in Ang II-induced apoptosis and the cardioprotective effect of Ghrelin. ( A ) MiR-208a negative control (NC), mimics and inhibitors with 5-carboxyfluorescein (5-FAM) were transfected into H9c2 cells by Fugene reagent. Whether the transfection is successful was detected using fluorescence microscope and confirmed by qRT-PCR. ( B, C ) After transfection for 24 hours, the cells were then treated with Ang II or Ang II+Ghrelin for another 24 hours. The percentage of apoptotic cells was measured by flow cytometry. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Ghrelin Alleviates Angiotensin II-Induced H9c2 Apoptosis: Impact of the miR-208 Family

    doi: 10.12659/MSM.908096

    Figure Lengend Snippet: The role of miR-208a in Ang II-induced apoptosis and the cardioprotective effect of Ghrelin. ( A ) MiR-208a negative control (NC), mimics and inhibitors with 5-carboxyfluorescein (5-FAM) were transfected into H9c2 cells by Fugene reagent. Whether the transfection is successful was detected using fluorescence microscope and confirmed by qRT-PCR. ( B, C ) After transfection for 24 hours, the cells were then treated with Ang II or Ang II+Ghrelin for another 24 hours. The percentage of apoptotic cells was measured by flow cytometry. * P

    Article Snippet: The miRNA mimics, inhibitors, and negative control (NC) were synthesized and labeled with 5-carboxyfluorescein (5-FAM) by Gene Pharma (Shanghai, China), and FuGENE HD reagents used for transfection were purchased from Promega (Madison, WI, USA).

    Techniques: Negative Control, Transfection, Fluorescence, Microscopy, Quantitative RT-PCR, Flow Cytometry, Cytometry

    Rb phosphorylation at S807 is required for association with Bax. Rb mutant plasmids were generated as described in the Materials and Methods section. Rb-negative C33A cells were transfected using Fugene (Promega). ( A ) Rb plasmids expressing either S807A or S811A alanine mutants were transfected into cells and 48 hours later cell lysates were utilized in GST-Bax fusion protein pull down assays. Proteins associated with GST-Bax were analyzed by immunoblotting with Rb antibodies. ( B ) Rb plasmids expressing either S807A or S807E mutants were transfected into C33A cells and 48 hours later co-immunoprecipitation with Bax antibodies was performed. Immunoprecipitates were analyzed by immunoblotting with Rb and Bax antibodies. Equivalent expression of the Rb mutant proteins in the C33A cells is verified by immunoblotting (lower panel). Data shown is representative of three independent experiments.

    Journal: Cell Cycle

    Article Title: Phosphorylation of the Retinoblastoma protein (Rb) on serine-807 is required for association with Bax

    doi: 10.4161/15384101.2014.964093

    Figure Lengend Snippet: Rb phosphorylation at S807 is required for association with Bax. Rb mutant plasmids were generated as described in the Materials and Methods section. Rb-negative C33A cells were transfected using Fugene (Promega). ( A ) Rb plasmids expressing either S807A or S811A alanine mutants were transfected into cells and 48 hours later cell lysates were utilized in GST-Bax fusion protein pull down assays. Proteins associated with GST-Bax were analyzed by immunoblotting with Rb antibodies. ( B ) Rb plasmids expressing either S807A or S807E mutants were transfected into C33A cells and 48 hours later co-immunoprecipitation with Bax antibodies was performed. Immunoprecipitates were analyzed by immunoblotting with Rb and Bax antibodies. Equivalent expression of the Rb mutant proteins in the C33A cells is verified by immunoblotting (lower panel). Data shown is representative of three independent experiments.

    Article Snippet: Plasmids were introduced into C33A cells using Fugene (Promega) performed according to the manufacturer's directions.

    Techniques: Mutagenesis, Generated, Transfection, Expressing, Immunoprecipitation

    The role of miR-208a in Ang II-induced apoptosis and the cardioprotective effect of Ghrelin. ( A ) MiR-208a negative control (NC), mimics and inhibitors with 5-carboxyfluorescein (5-FAM) were transfected into H9c2 cells by Fugene reagent. Whether the transfection is successful was detected using fluorescence microscope and confirmed by qRT-PCR. ( B, C ) After transfection for 24 hours, the cells were then treated with Ang II or Ang II+Ghrelin for another 24 hours. The percentage of apoptotic cells was measured by flow cytometry. * P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Ghrelin Alleviates Angiotensin II-Induced H9c2 Apoptosis: Impact of the miR-208 Family

    doi: 10.12659/MSM.908096

    Figure Lengend Snippet: The role of miR-208a in Ang II-induced apoptosis and the cardioprotective effect of Ghrelin. ( A ) MiR-208a negative control (NC), mimics and inhibitors with 5-carboxyfluorescein (5-FAM) were transfected into H9c2 cells by Fugene reagent. Whether the transfection is successful was detected using fluorescence microscope and confirmed by qRT-PCR. ( B, C ) After transfection for 24 hours, the cells were then treated with Ang II or Ang II+Ghrelin for another 24 hours. The percentage of apoptotic cells was measured by flow cytometry. * P

    Article Snippet: The FuGENE transfection reagent was purchased from Promega (Madison, WI, USA).

    Techniques: Negative Control, Transfection, Fluorescence, Microscopy, Quantitative RT-PCR, Flow Cytometry, Cytometry

    Analysis of cellular protein expression in ZIKV-infected hBMECs. (A to C) hBMECs were mock infected or infected with ZIKV, and 1 to 9 dpi supernatants were analyzed in an ELISA (R D Systems) for CCL/RANTES (A), IFN-β (B), and IFN-λ (C) levels relative to antigen standards. As an hBMEC IFN-β response control, we transfected hBMECs with poly(I/C) (1 µg/ml) and Fugene6 at 3:1 and evaluated secreted IFN-β levels in supernatants via ELISA (36 h posttransfection) (D) Western blot analysis of MXA and IFIT1 genes, and GAPDH controls, in lysates from mock-infected or ZIKV-infected hBMECs (1 to 9 dpi). IFIT and MxA protein levels in ZIKV-infected hBMECs 9 dpi versus results 6 h post-IFN-α treatment (1,000 U/ml).

    Journal: mBio

    Article Title: Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells

    doi: 10.1128/mBio.00952-17

    Figure Lengend Snippet: Analysis of cellular protein expression in ZIKV-infected hBMECs. (A to C) hBMECs were mock infected or infected with ZIKV, and 1 to 9 dpi supernatants were analyzed in an ELISA (R D Systems) for CCL/RANTES (A), IFN-β (B), and IFN-λ (C) levels relative to antigen standards. As an hBMEC IFN-β response control, we transfected hBMECs with poly(I/C) (1 µg/ml) and Fugene6 at 3:1 and evaluated secreted IFN-β levels in supernatants via ELISA (36 h posttransfection) (D) Western blot analysis of MXA and IFIT1 genes, and GAPDH controls, in lysates from mock-infected or ZIKV-infected hBMECs (1 to 9 dpi). IFIT and MxA protein levels in ZIKV-infected hBMECs 9 dpi versus results 6 h post-IFN-α treatment (1,000 U/ml).

    Article Snippet: Levels of IFN-β, IFN-λ, and CCL5/RANTES in the supernatants of mock- and ZIKV-infected hBMECs at 12 hpi to 9 dpi were measured using a DuoSet ELISA (R & D Systems). hBMECs transfected with poly(I/C) (1 µg/ml) and Fugene6 (3:1; Promega) were similarly evaluated for secreted IFN-β at 36 h posttransfection.

    Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Western Blot

    CHO cells transfected with Sra did not display any LPS binding. CHO cells were transfected with murine Sra (plasmid) using Fugene HD. After 24 h of the transfection, cells were analyzed using various reagents. (A) Mock-transfected CHO cell; (B–E)

    Journal: Innate immunity

    Article Title: Deletion of scavenger receptor A gene in mice resulted in protection from septic shock and modulation of TLR4 signaling in isolated peritoneal macrophages

    doi: 10.1177/1753425912449548

    Figure Lengend Snippet: CHO cells transfected with Sra did not display any LPS binding. CHO cells were transfected with murine Sra (plasmid) using Fugene HD. After 24 h of the transfection, cells were analyzed using various reagents. (A) Mock-transfected CHO cell; (B–E)

    Article Snippet: CHO-K1 cells (5×105 cells) were transfected with Sra gene in pCMVscript vector (2 μg of DNA) using fugene reagent (2:1 reagent/DNA ratio; Promega, Madison, WI).

    Techniques: Transfection, Binding Assay, Plasmid Preparation

    CHO cells transfected with SraI, SraII, or both did not bind LPS. CHO cells were transfected with murine SraI, SraII, or both (plasmid) using Fugene HD. After 24 h of the transfection, cells were analyzed using various reagents. SraI -transfected CHO cells

    Journal: Innate immunity

    Article Title: Deletion of scavenger receptor A gene in mice resulted in protection from septic shock and modulation of TLR4 signaling in isolated peritoneal macrophages

    doi: 10.1177/1753425912449548

    Figure Lengend Snippet: CHO cells transfected with SraI, SraII, or both did not bind LPS. CHO cells were transfected with murine SraI, SraII, or both (plasmid) using Fugene HD. After 24 h of the transfection, cells were analyzed using various reagents. SraI -transfected CHO cells

    Article Snippet: CHO-K1 cells (5×105 cells) were transfected with Sra gene in pCMVscript vector (2 μg of DNA) using fugene reagent (2:1 reagent/DNA ratio; Promega, Madison, WI).

    Techniques: Transfection, Plasmid Preparation

    ChIP analysis indicating that ZPO2 and ZBTB32 occupy the Gata3 promoter. EpH4.9 cells were grown in 150-mm culture dishes and cotransfected with full-length Zpo2 and Myc-tagged ZBTB32 (generous gifts from Dr. I.-Cheng Ho, Harvard Medical School) constructs via FuGENE6 transfection reagent (Promega). ChIP analysis was performed using the ChIP-IT High-Sensitivity Kit (Active Motif) in accordance with the manufacturer’s protocol. qPCR for ChIP samples was performed using Gata3 promoter sequence primers listed in the text. ChIP analysis indicated that both ZPO2 and ZBTB32 occupy the promoter sequence of GATA3.

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

    Article Title: ZNF503/Zpo2 drives aggressive breast cancer progression by down-regulation of GATA3 expression

    doi: 10.1073/pnas.1701690114

    Figure Lengend Snippet: ChIP analysis indicating that ZPO2 and ZBTB32 occupy the Gata3 promoter. EpH4.9 cells were grown in 150-mm culture dishes and cotransfected with full-length Zpo2 and Myc-tagged ZBTB32 (generous gifts from Dr. I.-Cheng Ho, Harvard Medical School) constructs via FuGENE6 transfection reagent (Promega). ChIP analysis was performed using the ChIP-IT High-Sensitivity Kit (Active Motif) in accordance with the manufacturer’s protocol. qPCR for ChIP samples was performed using Gata3 promoter sequence primers listed in the text. ChIP analysis indicated that both ZPO2 and ZBTB32 occupy the promoter sequence of GATA3.

    Article Snippet: EpH4.9 cells were cultured in 10-cm culture dishes and cotransfected with 7 µg of V5-tagged full-length Zpo2 and 7 µg of Myc-tagged ZBTB32 using FuGENE6 transfection reagent (Promega).

    Techniques: Chromatin Immunoprecipitation, Construct, Transfection, Real-time Polymerase Chain Reaction, Sequencing

    MafB-deficient T-ALL cells downregulate Notch targets and perform poorly in competitive culture conditions A. MafB expression measured by qRT-PCR in T6E cells 48 hours after treatment with indicated siRNAs relative to that in cells cultured with transfection reagent (untreated). * p≤0.05, ** p≤0.005 by unpaired t-test. B. Relative expression analysis by qRT-PCR of Notch signaling and Myc signaling (CAD) targets and expression controls (GAPDH) in MafB-siRNA#1–treated cells relative to each in Scrmb-siRNA–treated cells. C. Chromatin immunoprecipitation (ChIP) assay measuring occupancy of P300 at the Hes1 promoter and at the Notch-dependent Myc enhancer (NDME) in MafB-shRNA#3–treated T6E cells. Genomic DNA sequence of GAPDH is used as non-specific control. ***p

    Journal: Science signaling

    Article Title: MafB is a novel enhancer of oncogenic Notch signaling in T-cell acute lymphoblastic leukemia

    doi: 10.1126/scisignal.aam6846

    Figure Lengend Snippet: MafB-deficient T-ALL cells downregulate Notch targets and perform poorly in competitive culture conditions A. MafB expression measured by qRT-PCR in T6E cells 48 hours after treatment with indicated siRNAs relative to that in cells cultured with transfection reagent (untreated). * p≤0.05, ** p≤0.005 by unpaired t-test. B. Relative expression analysis by qRT-PCR of Notch signaling and Myc signaling (CAD) targets and expression controls (GAPDH) in MafB-siRNA#1–treated cells relative to each in Scrmb-siRNA–treated cells. C. Chromatin immunoprecipitation (ChIP) assay measuring occupancy of P300 at the Hes1 promoter and at the Notch-dependent Myc enhancer (NDME) in MafB-shRNA#3–treated T6E cells. Genomic DNA sequence of GAPDH is used as non-specific control. ***p

    Article Snippet: The Matrix Wellmate (Thermo) was used to dispense the transfection mix containing the reporter and Notch1 mutant plasmids in combination with transfection reagent (Fugene6 Promega), which were added to the wells (4000 U20S cells/well) after a 30 min incubation.

    Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Transfection, Chromatin Immunoprecipitation, shRNA, Sequencing

    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by FuGENE 6 Transfection reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.

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

    Article Title: Effects of mutant p53 expression on human 15-lipoxygenase-promoter activity and murine 12/15-lipoxygenase gene expression: Evidence that 15-lipoxygenase is a mutator gene

    doi:

    Figure Lengend Snippet: Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by FuGENE 6 Transfection reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.

    Article Snippet: After 24 h, the plasmid DNA (1 μg) was transfected into the cells by FuGENE 6 Transfection reagent (Boehringer Mannheim).

    Techniques: Luciferase, Construct, Sequencing, Transfection, Activity Assay

    Immunocytofluorescence detection of receptors in DU145 cells. DU145 cells were seeded on two-well Lab Tek Chamber slides (Nalge) 18 hr before transfection. One to two micrograms of DNA per 10 5 cells was transfected either with the AR (unliganded or liganded), TR4, or ER alone or in combination with the FuGENE6 transfection reagent (Boehringer-Mannheim). After 24 hr transfection, cells were treated with 100 nM DHT or ethanol. Immunostaining was performed by incubation with the anti-AR polyclonal antibody, anti-TR4 mAb, or anti-ERα mAb, followed by incubation with either fluorescein-conjugated goat anti-rabbit or anti-mouse antibodies (ICN). The red signal represents the AR, the green signal represents the TR4 or ER, and the yellow signal represents colocalization of the two signals in cotransfection of both receptors (the AR with TR4 or ER). Shown is the immunostaining of a single transfection of ( A ) unliganded AR, ( B ) 100 nM DHT treatment of the AR, ( C ) TR4, and ( D ) ER, and for cotransfection of ( E ) AR with TR4 and ( F ) AR with ER. The slides were photographed under 100-fold magnification by using confocal microscopy.

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

    Article Title: Convergence of two repressors through heterodimer formation of androgen receptor and testicular orphan receptor-4: A unique signaling pathway in the steroid receptor superfamily

    doi:

    Figure Lengend Snippet: Immunocytofluorescence detection of receptors in DU145 cells. DU145 cells were seeded on two-well Lab Tek Chamber slides (Nalge) 18 hr before transfection. One to two micrograms of DNA per 10 5 cells was transfected either with the AR (unliganded or liganded), TR4, or ER alone or in combination with the FuGENE6 transfection reagent (Boehringer-Mannheim). After 24 hr transfection, cells were treated with 100 nM DHT or ethanol. Immunostaining was performed by incubation with the anti-AR polyclonal antibody, anti-TR4 mAb, or anti-ERα mAb, followed by incubation with either fluorescein-conjugated goat anti-rabbit or anti-mouse antibodies (ICN). The red signal represents the AR, the green signal represents the TR4 or ER, and the yellow signal represents colocalization of the two signals in cotransfection of both receptors (the AR with TR4 or ER). Shown is the immunostaining of a single transfection of ( A ) unliganded AR, ( B ) 100 nM DHT treatment of the AR, ( C ) TR4, and ( D ) ER, and for cotransfection of ( E ) AR with TR4 and ( F ) AR with ER. The slides were photographed under 100-fold magnification by using confocal microscopy.

    Article Snippet: One or two micrograms of DNA per 105 cells was transfected by the FuGENE6 transfection reagent (Boehringer-Mannheim).

    Techniques: Transfection, Immunostaining, Incubation, Cotransfection, Confocal Microscopy