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TaKaRa pcmv βgal
Pcmv βgal, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 89 stars, based on 15 article reviews
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pcmv βgal - by Bioz Stars, 2020-09
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Clone Assay:

Article Title: Importin 7 and Importin ?/Importin ? Are Nuclear Import Receptors for the Glucocorticoid Receptor
Article Snippet: .. The lacZ gene was amplified from pCMV-βgal ( ) and cloned into the Bgl II site of pEGFP-C1 (CLONTECH, Palo Alto, CA) producing pEGFP- lacZ . .. A DNA fragment encoding amino acids 540–795 of GR was amplified from pEGFP-N795 with primers containing Sal I and Bgl II sites and cloned into the Sal I- Bam HI sites of pEGFP- lacZ . pJW248, containing the ADH1 promoter and GFP, was kindly provided by Jonathan Weissman (UCSF). pAdh-N795-GFP, wt or K513-515A, was constructed by excising full-length rat GR from pEGFP-N795 or pEGFP-N795 with Bam HI- Xho I and ligating into the Sal I- Bgl II sites of pJW248. pGPD-N795-GFP and pGPD-N795 K513-515A GFP (expressing GR from a GPD promoter) were created by excising GR-GFP from pAdh-N795-GFP or pAdh-N795-GFP-K513-515A with Bam HI- Bgl II and subcloning into the Bam HI site of pRS316-GPD-PGK, a kind gift from Anastasia Kralli (Scripps). pYN525-GFP and pYN525 K513-515A GFP were created by amplifying a DNA fragment encoding amino acids 1–525 from pEGFP-N795 and pEGFP-N795 K513-515A using primers containing Xho I and Bam HI restriction sites.

Amplification:

Article Title: Importin 7 and Importin ?/Importin ? Are Nuclear Import Receptors for the Glucocorticoid Receptor
Article Snippet: .. The lacZ gene was amplified from pCMV-βgal ( ) and cloned into the Bgl II site of pEGFP-C1 (CLONTECH, Palo Alto, CA) producing pEGFP- lacZ . .. A DNA fragment encoding amino acids 540–795 of GR was amplified from pEGFP-N795 with primers containing Sal I and Bgl II sites and cloned into the Sal I- Bam HI sites of pEGFP- lacZ . pJW248, containing the ADH1 promoter and GFP, was kindly provided by Jonathan Weissman (UCSF). pAdh-N795-GFP, wt or K513-515A, was constructed by excising full-length rat GR from pEGFP-N795 or pEGFP-N795 with Bam HI- Xho I and ligating into the Sal I- Bgl II sites of pJW248. pGPD-N795-GFP and pGPD-N795 K513-515A GFP (expressing GR from a GPD promoter) were created by excising GR-GFP from pAdh-N795-GFP or pAdh-N795-GFP-K513-515A with Bam HI- Bgl II and subcloning into the Bam HI site of pRS316-GPD-PGK, a kind gift from Anastasia Kralli (Scripps). pYN525-GFP and pYN525 K513-515A GFP were created by amplifying a DNA fragment encoding amino acids 1–525 from pEGFP-N795 and pEGFP-N795 K513-515A using primers containing Xho I and Bam HI restriction sites.

Plasmid Preparation:

Article Title: Tetracycline-Regulated Suppression of Amber Codons in Mammalian Cells
Article Snippet: .. The transcription of the histidine-tagged E. coli GlnRS gene is controlled by tTA in pUtetO-16GlnRS and is constitutive from the CMV enhancer/promoter in pc-16GlnRS. pCMV-βgal, containing an E. coli β-galactosidase gene transcribed by using the CMV promoter (Clontech), was used as a control vector. .. HeLa-tetOff cells (Clontech), which express constitutively tTA, were maintained at 37°C with 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% calf serum, 100 U of penicillin G sodium, 100 μg of streptomycin sulfate per ml, and 200 μg of G418 (GIBCO BRL) per ml.

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  • 89
    TaKaRa pcmv βgal
    ) plus pHOOK (3 μg) plus <t>pCMV-βGAL</t> (0.1 μg) with increasing amounts of KCREB (0.5, 1, 3, and 5 μg [A and B]), p300ΔHAT (0.5, 1, 3, and 5 μg [C and D]), or P/CAF NP (0.5, 1, 3, and 5 μg [E and F]). Total amounts of transfected DNA were normalized with pUC19. Each series of transfections was repeated three or more times. Luciferase activities were assayed 48 h after transfection. Luciferase activities in lanes 1 and 5 are significantly different at the indicated P values ( P
    Pcmv βgal, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcmv βgal/product/TaKaRa
    Average 89 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    pcmv βgal - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    88
    TaKaRa transfection control plasmid
    Deletion analysis of the 5′-flanking region of the Mrp2 gene The 5′-flanking region constructs were transiently transfected into Hepa 1-6 cells, and after 24 h the LUC ( A ) and CAT ( B ) activities were determined. The results are expressed as the percentage activity of the entire isolated 5′-flanking region (with p−1895/+99-LUC or p−1895/+99-CAT normalized to 100%), after subtraction of the activity of the empty plasmid reporter. <t>Transfections</t> were performed in triplicate, and results are means±S.D. ( C ) Schematic representation of the 5′-flanking region of the Mrp2 gene. The reporter gene constructs and the putative binding sites for transcription factors, including ARE - 1 and ARE-2 sequences, are shown. AP-1, activator protein 1.
    Transfection Control Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfection control plasmid/product/TaKaRa
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transfection control plasmid - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

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    ) plus pHOOK (3 μg) plus pCMV-βGAL (0.1 μg) with increasing amounts of KCREB (0.5, 1, 3, and 5 μg [A and B]), p300ΔHAT (0.5, 1, 3, and 5 μg [C and D]), or P/CAF NP (0.5, 1, 3, and 5 μg [E and F]). Total amounts of transfected DNA were normalized with pUC19. Each series of transfections was repeated three or more times. Luciferase activities were assayed 48 h after transfection. Luciferase activities in lanes 1 and 5 are significantly different at the indicated P values ( P

    Journal: Journal of Virology

    Article Title: Differential Requirements for Activation of Integrated and Transiently Transfected Human T-Cell Leukemia Virus Type 1 Long Terminal Repeat

    doi: 10.1128/JVI.76.24.12564-12573.2002

    Figure Lengend Snippet: ) plus pHOOK (3 μg) plus pCMV-βGAL (0.1 μg) with increasing amounts of KCREB (0.5, 1, 3, and 5 μg [A and B]), p300ΔHAT (0.5, 1, 3, and 5 μg [C and D]), or P/CAF NP (0.5, 1, 3, and 5 μg [E and F]). Total amounts of transfected DNA were normalized with pUC19. Each series of transfections was repeated three or more times. Luciferase activities were assayed 48 h after transfection. Luciferase activities in lanes 1 and 5 are significantly different at the indicated P values ( P

    Article Snippet: pHpX , pRSV-neo , and the HTLV-1 LTR-luciferase (LTR-luc) plasmid pHTLV-luc ( ) were described previously. pHOOK-1 was obtained from Invitrogen (Carlsbad, Calif.), and pCMV-βGAL was obtained from Clontech (Palo Alto, Calif.). pCI-P/CAF deletion mutants were produced by digestion with the appropriate restriction enzymes followed by in-frame ligation.

    Techniques: Transfection, Luciferase

    Activation of transiently transfected and stably integrated HTLV-luc by CREB, p300, and P/CAF. Transfections were performed with CHOK1 (A, D, and G) and HeLa (B, E, and H) cells. To assess the expression of transiently transfected reporter plasmid (open bars), CHOK1 cells were transfected with 1 ng of pHTLV-luc, and HeLa cells were transfected with 10 ng of pHTLV-luc. Additionally, 0.1 μg of pCMV-βGAL (as a transfection control) and 0.2, 1, or 3 μg of CREB (A), or p300 (D), or P/CAF (G) were cotransfected as indicated. To assess the expression of integrated HTLV-1 LTR-luc (filled bars), CHOK1-luc or HeLa-luc cells were transfected with 3 μg of pHOOK plus 0.1 μg of pCMV-βGAL and 0.2, 1, or 3 μg of CREB (B), p300 (E), or P/CAF (H). Forty-eight hours after transfection, luciferase activities were measured. Luciferase values are averages ± standard deviations from three or more independent transfections. Statistical significance at the indicated P values was verified with the one-tailed t test. Panels C, F, and I show Western blotting of endogenous (pUC19 lanes) CREB (C), p300 (F), and P/CAF (I) in CHOK1 and HeLa cells or the total amounts of CREB, p300, and P/CAF after transfection of 3 μg of the respective expression plasmid.

    Journal: Journal of Virology

    Article Title: Differential Requirements for Activation of Integrated and Transiently Transfected Human T-Cell Leukemia Virus Type 1 Long Terminal Repeat

    doi: 10.1128/JVI.76.24.12564-12573.2002

    Figure Lengend Snippet: Activation of transiently transfected and stably integrated HTLV-luc by CREB, p300, and P/CAF. Transfections were performed with CHOK1 (A, D, and G) and HeLa (B, E, and H) cells. To assess the expression of transiently transfected reporter plasmid (open bars), CHOK1 cells were transfected with 1 ng of pHTLV-luc, and HeLa cells were transfected with 10 ng of pHTLV-luc. Additionally, 0.1 μg of pCMV-βGAL (as a transfection control) and 0.2, 1, or 3 μg of CREB (A), or p300 (D), or P/CAF (G) were cotransfected as indicated. To assess the expression of integrated HTLV-1 LTR-luc (filled bars), CHOK1-luc or HeLa-luc cells were transfected with 3 μg of pHOOK plus 0.1 μg of pCMV-βGAL and 0.2, 1, or 3 μg of CREB (B), p300 (E), or P/CAF (H). Forty-eight hours after transfection, luciferase activities were measured. Luciferase values are averages ± standard deviations from three or more independent transfections. Statistical significance at the indicated P values was verified with the one-tailed t test. Panels C, F, and I show Western blotting of endogenous (pUC19 lanes) CREB (C), p300 (F), and P/CAF (I) in CHOK1 and HeLa cells or the total amounts of CREB, p300, and P/CAF after transfection of 3 μg of the respective expression plasmid.

    Article Snippet: pHpX , pRSV-neo , and the HTLV-1 LTR-luciferase (LTR-luc) plasmid pHTLV-luc ( ) were described previously. pHOOK-1 was obtained from Invitrogen (Carlsbad, Calif.), and pCMV-βGAL was obtained from Clontech (Palo Alto, Calif.). pCI-P/CAF deletion mutants were produced by digestion with the appropriate restriction enzymes followed by in-frame ligation.

    Techniques: Activation Assay, Transfection, Stable Transfection, Expressing, Plasmid Preparation, Luciferase, One-tailed Test, Western Blot

    Southern blotting analyses of stably integrated HTLV-1 LTR-luc in CHOK1 and HeLa cells. (A) Three independently cloned CHOK1 and HeLa cells with a stably integrated HTLV-1 LTR-luc gene are shown. The autoradiographs compare luciferase-specific hybridzation signals from parental cell DNAs (CHOK1, lane 4; HeLa, lane 8) and DNAs from three independently selected CHOK1 (lanes 5 to 7) or HeLa (lanes 9 to 11) integrants. Lanes 1 to 3 show reconstruction controls in which 100, 10, or 1 copy of linearized pGL3-basic plasmid (4.8 kbp) was mixed with Eco RI-digested CV-1 genomic DNA. (B) Comparisons of luciferase activities from transiently transfected pHTLV-luc plasmid and stably integrated pHTLV-luc. One, 10, or 100 ng of pHTLV-luc was transiently transfected into CHOK1 or HeLa cells. Transiently transfected cells were compared to two pools of CHOK1 and HeLa integrants for luciferase activity as described in Materials and Methods. Absolute luciferase values were normalized to β-galactosidase activity from a cotransfected pCMV-βGAL plasmid.

    Journal: Journal of Virology

    Article Title: Differential Requirements for Activation of Integrated and Transiently Transfected Human T-Cell Leukemia Virus Type 1 Long Terminal Repeat

    doi: 10.1128/JVI.76.24.12564-12573.2002

    Figure Lengend Snippet: Southern blotting analyses of stably integrated HTLV-1 LTR-luc in CHOK1 and HeLa cells. (A) Three independently cloned CHOK1 and HeLa cells with a stably integrated HTLV-1 LTR-luc gene are shown. The autoradiographs compare luciferase-specific hybridzation signals from parental cell DNAs (CHOK1, lane 4; HeLa, lane 8) and DNAs from three independently selected CHOK1 (lanes 5 to 7) or HeLa (lanes 9 to 11) integrants. Lanes 1 to 3 show reconstruction controls in which 100, 10, or 1 copy of linearized pGL3-basic plasmid (4.8 kbp) was mixed with Eco RI-digested CV-1 genomic DNA. (B) Comparisons of luciferase activities from transiently transfected pHTLV-luc plasmid and stably integrated pHTLV-luc. One, 10, or 100 ng of pHTLV-luc was transiently transfected into CHOK1 or HeLa cells. Transiently transfected cells were compared to two pools of CHOK1 and HeLa integrants for luciferase activity as described in Materials and Methods. Absolute luciferase values were normalized to β-galactosidase activity from a cotransfected pCMV-βGAL plasmid.

    Article Snippet: pHpX , pRSV-neo , and the HTLV-1 LTR-luciferase (LTR-luc) plasmid pHTLV-luc ( ) were described previously. pHOOK-1 was obtained from Invitrogen (Carlsbad, Calif.), and pCMV-βGAL was obtained from Clontech (Palo Alto, Calif.). pCI-P/CAF deletion mutants were produced by digestion with the appropriate restriction enzymes followed by in-frame ligation.

    Techniques: Southern Blot, Stable Transfection, Clone Assay, Luciferase, Plasmid Preparation, Transfection, Activity Assay

    Definition of regions within P/CAF that contribute to the activation of the HTLV-1 LTR. (A) Schematic depictions of six P/CAF deletion mutants: ΔHAT, BA, NA, NB, NP, and NE. The full-length Flag-tagged P/CAF protein is shown at top with the HAT and BROMO domains indicated. (B) Efficient expression of transfected P/CAF and P/CAF mutants in CHOK1 and HeLa cells. Cells were transfected with 3 μg of Flag-tagged pCI-P/CAF or the indicated deletion mutant followed by Western blotting with anti-Flag. The NA mutant, due to its small size, could not be detected on this gel. The BA mutant was not assayed in this experiment. (C) Activation of integrated HTLV-luc by P/CAF and P/CAF mutants. Luciferase values are averages ± standard deviations from three independent transfections of pooled CHOK1-luc cells with the indicated P/CAF plasmids. (D and E) Dominant-negative activity of P/CAF NP. Pooled CHOK1-luc cells were transfected with pHOOK (3 μg) plus pCMV-βGAL (0.1 μg) plus pCI-P/CAF (0.5 μg) with increasing amounts of P/CAF NP (0.5, 1, and 3 μg), either with (D) or without (E) Tax. The total amounts of transfected plasmid were normalized with pUC19. Luciferase values are averages ± standard deviations from three independent transfections. A significant difference in luciferase activities for lanes 3 and 6 was seen ( P

    Journal: Journal of Virology

    Article Title: Differential Requirements for Activation of Integrated and Transiently Transfected Human T-Cell Leukemia Virus Type 1 Long Terminal Repeat

    doi: 10.1128/JVI.76.24.12564-12573.2002

    Figure Lengend Snippet: Definition of regions within P/CAF that contribute to the activation of the HTLV-1 LTR. (A) Schematic depictions of six P/CAF deletion mutants: ΔHAT, BA, NA, NB, NP, and NE. The full-length Flag-tagged P/CAF protein is shown at top with the HAT and BROMO domains indicated. (B) Efficient expression of transfected P/CAF and P/CAF mutants in CHOK1 and HeLa cells. Cells were transfected with 3 μg of Flag-tagged pCI-P/CAF or the indicated deletion mutant followed by Western blotting with anti-Flag. The NA mutant, due to its small size, could not be detected on this gel. The BA mutant was not assayed in this experiment. (C) Activation of integrated HTLV-luc by P/CAF and P/CAF mutants. Luciferase values are averages ± standard deviations from three independent transfections of pooled CHOK1-luc cells with the indicated P/CAF plasmids. (D and E) Dominant-negative activity of P/CAF NP. Pooled CHOK1-luc cells were transfected with pHOOK (3 μg) plus pCMV-βGAL (0.1 μg) plus pCI-P/CAF (0.5 μg) with increasing amounts of P/CAF NP (0.5, 1, and 3 μg), either with (D) or without (E) Tax. The total amounts of transfected plasmid were normalized with pUC19. Luciferase values are averages ± standard deviations from three independent transfections. A significant difference in luciferase activities for lanes 3 and 6 was seen ( P

    Article Snippet: pHpX , pRSV-neo , and the HTLV-1 LTR-luciferase (LTR-luc) plasmid pHTLV-luc ( ) were described previously. pHOOK-1 was obtained from Invitrogen (Carlsbad, Calif.), and pCMV-βGAL was obtained from Clontech (Palo Alto, Calif.). pCI-P/CAF deletion mutants were produced by digestion with the appropriate restriction enzymes followed by in-frame ligation.

    Techniques: Activation Assay, HAT Assay, Expressing, Transfection, Mutagenesis, Western Blot, Luciferase, Dominant Negative Mutation, Activity Assay, Plasmid Preparation

    NPR1/GCA promoter activity is inhibited by GREBP overexpression, and this inhibition is dependent on cGMP-RE. A , HEK293 cells were co-transfected with 10 ng of pCMV-βgal, 200 ng of hGCAp-pGL3b, and a combination of varying amounts of pCDNA1-Neo and/or pCDNA1-Neo-GREBP for a total plasmid amount of 500 ng/test. GREBP overexpression was confirmed by sqRT-PCR with specific primers targeting GREBP expressed by pCDNA1-Neo-GREBP. B , HEK293 cells were co-transfected with 10 ng of pCMVβ, 400 ng of pCDNA1-Neo or pCDNA1-Neo-GREBP, and 200 ng of hGCAp-pGL3b or the deleted cGMP-RE form of the human NPR1/GCA promoter hGCAp(ΔcGMP-RE)-pGL3b. All values are expressed as means ± S.E. ( error bars ) of relative luciferase activity from four different experiments performed in triplicate ( n = 12). *, p

    Journal: The Journal of Biological Chemistry

    Article Title: GREBP, a cGMP-response Element-binding Protein Repressing the Transcription of Natriuretic Peptide Receptor 1 (NPR1/GCA) *

    doi: 10.1074/jbc.M109.061622

    Figure Lengend Snippet: NPR1/GCA promoter activity is inhibited by GREBP overexpression, and this inhibition is dependent on cGMP-RE. A , HEK293 cells were co-transfected with 10 ng of pCMV-βgal, 200 ng of hGCAp-pGL3b, and a combination of varying amounts of pCDNA1-Neo and/or pCDNA1-Neo-GREBP for a total plasmid amount of 500 ng/test. GREBP overexpression was confirmed by sqRT-PCR with specific primers targeting GREBP expressed by pCDNA1-Neo-GREBP. B , HEK293 cells were co-transfected with 10 ng of pCMVβ, 400 ng of pCDNA1-Neo or pCDNA1-Neo-GREBP, and 200 ng of hGCAp-pGL3b or the deleted cGMP-RE form of the human NPR1/GCA promoter hGCAp(ΔcGMP-RE)-pGL3b. All values are expressed as means ± S.E. ( error bars ) of relative luciferase activity from four different experiments performed in triplicate ( n = 12). *, p

    Article Snippet: The day before transfection, HEK293 cells were seeded in a 12-well plate (7 × 104 cells/well), and plasmids hGCAp-pGL3p or hGCAp(ΔcGMP-RE)-pGL3b, pCDNA1-Neo, and/or pCDNA1-Neo-GREBP (for overexpression experiments), pSilencer 2.0-U6-NT or pSilencer 2.0-U6-shRNA-GREBP (for silencing experiments), and internal control pCMV-βgal (Clontech) plasmids were transfected and allowed to grow for 24 h. The cells were then lysed with Promega reporter buffer.

    Techniques: Activity Assay, Over Expression, Inhibition, Transfection, Plasmid Preparation, Polymerase Chain Reaction, Luciferase

    Effect of immunization protocol on T cells cytokines secretion of Der f mice (open bar, n = 6), pCMV-βgal (grey bar, n = 7) and pVAX-Der f1 (black bar, n = 7) when using 10 µg of DNA. One day after the last airway allergen challenge, mice were sacrificed and lung cells were cultured. The concentration of IL-4, IL-5, IL-13, IFN-γ, IL-10 and IL-17 were measured by flow cytometry. Results are expressed as the mean and standard deviation for each group. *p

    Journal: PLoS ONE

    Article Title: Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma

    doi: 10.1371/journal.pone.0085976

    Figure Lengend Snippet: Effect of immunization protocol on T cells cytokines secretion of Der f mice (open bar, n = 6), pCMV-βgal (grey bar, n = 7) and pVAX-Der f1 (black bar, n = 7) when using 10 µg of DNA. One day after the last airway allergen challenge, mice were sacrificed and lung cells were cultured. The concentration of IL-4, IL-5, IL-13, IFN-γ, IL-10 and IL-17 were measured by flow cytometry. Results are expressed as the mean and standard deviation for each group. *p

    Article Snippet: Plasmid Preparation and Formulation The pCMV-βgal ( = pCMV-LacZ) plasmid (Clontech, St Germain en Laye, France) encoding β-galactosidase, and the pVAX-Der f1 plasmid encoding Der f1, under the control of the human cytomegalovirus immediate promoter was used.

    Techniques: Mouse Assay, Cell Culture, Concentration Assay, Flow Cytometry, Cytometry, Standard Deviation

    Immunization protocols against Der f1 in asthmatic mice. pVAX-Der f1 or pCMV-βgal plasmid were injected i.m at days −28 and −7. Mice were then epicutaneously sensitized and intranasally challenged with total extract of HDM. Analyses were performed on day 35.

    Journal: PLoS ONE

    Article Title: Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma

    doi: 10.1371/journal.pone.0085976

    Figure Lengend Snippet: Immunization protocols against Der f1 in asthmatic mice. pVAX-Der f1 or pCMV-βgal plasmid were injected i.m at days −28 and −7. Mice were then epicutaneously sensitized and intranasally challenged with total extract of HDM. Analyses were performed on day 35.

    Article Snippet: Plasmid Preparation and Formulation The pCMV-βgal ( = pCMV-LacZ) plasmid (Clontech, St Germain en Laye, France) encoding β-galactosidase, and the pVAX-Der f1 plasmid encoding Der f1, under the control of the human cytomegalovirus immediate promoter was used.

    Techniques: Mouse Assay, Plasmid Preparation, Injection

    Effect of immunization protocol on the immune response of asthmatic mice. A. Splenocytes were stimulated overnight with a pool of Der f1 immunodominant peptides. The number of IFN-γ SFCs was determined. Der f (n = 2), pCMV-βgal (n = 5), pVAX-Der f1 (n = 6). B. Lung cells were stimulated overnight with a pool of Der f1 immunodominant peptides. n = 3 mice per group. The number of IFNγ spot forming colonies (SFCs) was determined. C. Humoral response was measured in sera one day after the last challenge in Der f (n = 6), pCMV-βgal (n = 7) and pVAX-Der f1 (n = 7) mice and additionally in control mice for IgE. Results are expressed as IgE, IgG1, IgG2a antibody titer. The mean number and standard deviation are shown for each group. *p

    Journal: PLoS ONE

    Article Title: Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma

    doi: 10.1371/journal.pone.0085976

    Figure Lengend Snippet: Effect of immunization protocol on the immune response of asthmatic mice. A. Splenocytes were stimulated overnight with a pool of Der f1 immunodominant peptides. The number of IFN-γ SFCs was determined. Der f (n = 2), pCMV-βgal (n = 5), pVAX-Der f1 (n = 6). B. Lung cells were stimulated overnight with a pool of Der f1 immunodominant peptides. n = 3 mice per group. The number of IFNγ spot forming colonies (SFCs) was determined. C. Humoral response was measured in sera one day after the last challenge in Der f (n = 6), pCMV-βgal (n = 7) and pVAX-Der f1 (n = 7) mice and additionally in control mice for IgE. Results are expressed as IgE, IgG1, IgG2a antibody titer. The mean number and standard deviation are shown for each group. *p

    Article Snippet: Plasmid Preparation and Formulation The pCMV-βgal ( = pCMV-LacZ) plasmid (Clontech, St Germain en Laye, France) encoding β-galactosidase, and the pVAX-Der f1 plasmid encoding Der f1, under the control of the human cytomegalovirus immediate promoter was used.

    Techniques: Mouse Assay, Standard Deviation

    Effect of prophylactic immunization protocol with 10 µg of Der f1 DNA on respiratory function. Airway resistance and Compliance was measured at day 35 using Flexivent with instillation of 5 to 20/ml methacholine in non asthmatic non vaccinated mice (n = 6, ) Der f (n = 7, O ), pCMV-βgal mice (n = 9, ) and pVAX-Der f1 (n = 9,▾) mice. Results are expressed in increased fold, as a mean for each group ± standard deviation. *p

    Journal: PLoS ONE

    Article Title: Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma

    doi: 10.1371/journal.pone.0085976

    Figure Lengend Snippet: Effect of prophylactic immunization protocol with 10 µg of Der f1 DNA on respiratory function. Airway resistance and Compliance was measured at day 35 using Flexivent with instillation of 5 to 20/ml methacholine in non asthmatic non vaccinated mice (n = 6, ) Der f (n = 7, O ), pCMV-βgal mice (n = 9, ) and pVAX-Der f1 (n = 9,▾) mice. Results are expressed in increased fold, as a mean for each group ± standard deviation. *p

    Article Snippet: Plasmid Preparation and Formulation The pCMV-βgal ( = pCMV-LacZ) plasmid (Clontech, St Germain en Laye, France) encoding β-galactosidase, and the pVAX-Der f1 plasmid encoding Der f1, under the control of the human cytomegalovirus immediate promoter was used.

    Techniques: Mouse Assay, Standard Deviation

    Effect of immunization protocol on BAL inflammation of non asthmatic non vaccinated mice (light grey bar, n = 8), Der f mice (open bar, n = 9), pCMV-βgal (grey bar, n = 7) and pVAX-Der f1 mice (black bar, n = 7) when using 10 µg of DNA. The total number of cells was determined by cell count on Kova slides. The cellular composition was established by flow cytometry. Results are expressed as absolute number of cells, as the mean and standard deviation for each group. **p

    Journal: PLoS ONE

    Article Title: Block Copolymer/DNA Vaccination Induces a Strong Allergen-Specific Local Response in a Mouse Model of House Dust Mite Asthma

    doi: 10.1371/journal.pone.0085976

    Figure Lengend Snippet: Effect of immunization protocol on BAL inflammation of non asthmatic non vaccinated mice (light grey bar, n = 8), Der f mice (open bar, n = 9), pCMV-βgal (grey bar, n = 7) and pVAX-Der f1 mice (black bar, n = 7) when using 10 µg of DNA. The total number of cells was determined by cell count on Kova slides. The cellular composition was established by flow cytometry. Results are expressed as absolute number of cells, as the mean and standard deviation for each group. **p

    Article Snippet: Plasmid Preparation and Formulation The pCMV-βgal ( = pCMV-LacZ) plasmid (Clontech, St Germain en Laye, France) encoding β-galactosidase, and the pVAX-Der f1 plasmid encoding Der f1, under the control of the human cytomegalovirus immediate promoter was used.

    Techniques: Mouse Assay, Cell Counting, Flow Cytometry, Cytometry, Standard Deviation

    Deletion analysis of the 5′-flanking region of the Mrp2 gene The 5′-flanking region constructs were transiently transfected into Hepa 1-6 cells, and after 24 h the LUC ( A ) and CAT ( B ) activities were determined. The results are expressed as the percentage activity of the entire isolated 5′-flanking region (with p−1895/+99-LUC or p−1895/+99-CAT normalized to 100%), after subtraction of the activity of the empty plasmid reporter. Transfections were performed in triplicate, and results are means±S.D. ( C ) Schematic representation of the 5′-flanking region of the Mrp2 gene. The reporter gene constructs and the putative binding sites for transcription factors, including ARE - 1 and ARE-2 sequences, are shown. AP-1, activator protein 1.

    Journal: Biochemical Journal

    Article Title: Role of Nrf2 in the regulation of the Mrp2 (ABCC2) gene

    doi: 10.1042/BJ20051518

    Figure Lengend Snippet: Deletion analysis of the 5′-flanking region of the Mrp2 gene The 5′-flanking region constructs were transiently transfected into Hepa 1-6 cells, and after 24 h the LUC ( A ) and CAT ( B ) activities were determined. The results are expressed as the percentage activity of the entire isolated 5′-flanking region (with p−1895/+99-LUC or p−1895/+99-CAT normalized to 100%), after subtraction of the activity of the empty plasmid reporter. Transfections were performed in triplicate, and results are means±S.D. ( C ) Schematic representation of the 5′-flanking region of the Mrp2 gene. The reporter gene constructs and the putative binding sites for transcription factors, including ARE - 1 and ARE-2 sequences, are shown. AP-1, activator protein 1.

    Article Snippet: The total DNA included 0.5 μg of reporter plasmid (p-insert-LUC or p-insert-CAT) and 0.1 μg of transfection control plasmid [pCMV-βgal (Clontech) or pRL-TK Renilla (Promega)] with or without 2 μg of the expression plasmid (p-Nrf2, p-Nrf2-DN or p-Nrf2-TA).

    Techniques: Construct, Transfection, Activity Assay, Isolation, Plasmid Preparation, Binding Assay