pcmv-lacz plasmid Search Results


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  • 88
    Promega pcmv lacz plasmid
    Pcmv Lacz Plasmid, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcmv lacz plasmid
    ORC1 binds SUV39H1 to control Cyclin E gene transcription. ORC1, but not ORC3 or ORC4 can repress gene transcription. ( A ) The U2OS cells were transfected with a Gal4-driven luciferase reporter as shown in the schematic with increasing amounts of Gal4DBD-ORC1 or Gal4DBD-SUV39H1 together with <t>pCMV-LacZ</t> plasmids. Relative luciferase activity was determined and normalized to lacZ activity. Experiments were carried out in triplicate. The whole cell extract was immunoblotted with anti-Gal4 antibody for expression of Gal4DBD fusion plasmids. α-Tubulin served as a loading control. Statistical analysis was performed using the Student’s t test. **p
    Pcmv Lacz Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Stratagene plasmid pcmv lacz
    ORC1 binds SUV39H1 to control Cyclin E gene transcription. ORC1, but not ORC3 or ORC4 can repress gene transcription. ( A ) The U2OS cells were transfected with a Gal4-driven luciferase reporter as shown in the schematic with increasing amounts of Gal4DBD-ORC1 or Gal4DBD-SUV39H1 together with <t>pCMV-LacZ</t> plasmids. Relative luciferase activity was determined and normalized to lacZ activity. Experiments were carried out in triplicate. The whole cell extract was immunoblotted with anti-Gal4 antibody for expression of Gal4DBD fusion plasmids. α-Tubulin served as a loading control. Statistical analysis was performed using the Student’s t test. **p
    Plasmid Pcmv Lacz, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    PlasmidFactory pcmv lacz plasmid
    Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with <t>pCMV</t> backbone, pCMV <t>lacZ,</t> enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P
    Pcmv Lacz Plasmid, supplied by PlasmidFactory, used in various techniques. Bioz Stars score: 88/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher β galactosidase expression plasmid pcmv lacz
    Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with <t>pCMV</t> backbone, pCMV <t>lacZ,</t> enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P
    β Galactosidase Expression Plasmid Pcmv Lacz, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Addgene inc pcmv gfp frt lacz plasmid
    Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with <t>pCMV</t> backbone, pCMV <t>lacZ,</t> enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P
    Pcmv Gfp Frt Lacz Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    TaKaRa beta galactosidase β gal reporter plasmid pcmv lacz
    Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with <t>pCMV</t> backbone, pCMV <t>lacZ,</t> enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P
    Beta Galactosidase β Gal Reporter Plasmid Pcmv Lacz, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa pcmv lacz
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    Pcmv Lacz, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pcmv lacz
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    Pcmv Lacz, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega pcmv lacz
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    Pcmv Lacz, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher expression vector pcmv lacz
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    Expression Vector Pcmv Lacz, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa β galactosidase reporter plasmid
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    β Galactosidase Reporter Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega β galactosidase expression plasmid
    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from <t>pCMV-lacZ.</t> Each bar on the graph represents the average of four separate transfections.
    β Galactosidase Expression Plasmid, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa pcmv βgal
    Effect of immunization protocol on T cells cytokines secretion of Der f mice (open bar, n = 6), <t>pCMV-βgal</t> (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
    Pcmv βgal, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 139 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa lacz expression vector
    Fusion assay between ASCT2 -transduced and <t>syncytin-Ory1</t> -transduced co-cultured cells demonstrates that ASCT2 is the syncytin-Ory1 receptor . Left panel: Cell-cell fusion was assayed upon independent transfections of a set of A23 cells with an empty vector (none) or an expression vector for either the syncytin-Ory1, syncytin-1 or syncytin-2 protein together with an nls- <t>LacZ</t> gene-expression vector, and another set of A23 cells with an expression vector for the syncytin-1 receptor ASCT2, the syncytin-2 receptor MFSD2 [ 13 ] or an empty vector (none). One day after transfection, cells were resuspended and pairs of transfected cells from each set were cocultured for 1-2 days, fixed and X-Gal stained. Right panel: Syncytia can be easily detected (arrows) for the syncytin-Ory1/ASCT2, syncytin-1/ASCT2 and syncytin-2/MFSD2 pairs, with only mononucleated cells visible in the other cases. Abbreviations: syn-Ory1, syncytin-Ory1; syn1, syncytin-1; syn2, syncytin-2.
    Lacz Expression Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Addgene inc human cell culture recombination assay pcmv gfp
    Fusion assay between ASCT2 -transduced and <t>syncytin-Ory1</t> -transduced co-cultured cells demonstrates that ASCT2 is the syncytin-Ory1 receptor . Left panel: Cell-cell fusion was assayed upon independent transfections of a set of A23 cells with an empty vector (none) or an expression vector for either the syncytin-Ory1, syncytin-1 or syncytin-2 protein together with an nls- <t>LacZ</t> gene-expression vector, and another set of A23 cells with an expression vector for the syncytin-1 receptor ASCT2, the syncytin-2 receptor MFSD2 [ 13 ] or an empty vector (none). One day after transfection, cells were resuspended and pairs of transfected cells from each set were cocultured for 1-2 days, fixed and X-Gal stained. Right panel: Syncytia can be easily detected (arrows) for the syncytin-Ory1/ASCT2, syncytin-1/ASCT2 and syncytin-2/MFSD2 pairs, with only mononucleated cells visible in the other cases. Abbreviations: syn-Ory1, syncytin-Ory1; syn1, syncytin-1; syn2, syncytin-2.
    Human Cell Culture Recombination Assay Pcmv Gfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ORC1 binds SUV39H1 to control Cyclin E gene transcription. ORC1, but not ORC3 or ORC4 can repress gene transcription. ( A ) The U2OS cells were transfected with a Gal4-driven luciferase reporter as shown in the schematic with increasing amounts of Gal4DBD-ORC1 or Gal4DBD-SUV39H1 together with pCMV-LacZ plasmids. Relative luciferase activity was determined and normalized to lacZ activity. Experiments were carried out in triplicate. The whole cell extract was immunoblotted with anti-Gal4 antibody for expression of Gal4DBD fusion plasmids. α-Tubulin served as a loading control. Statistical analysis was performed using the Student’s t test. **p

    Journal: eLife

    Article Title: Opposing roles for DNA replication initiator proteins ORC1 and CDC6 in control of Cyclin E gene transcription

    doi: 10.7554/eLife.12785

    Figure Lengend Snippet: ORC1 binds SUV39H1 to control Cyclin E gene transcription. ORC1, but not ORC3 or ORC4 can repress gene transcription. ( A ) The U2OS cells were transfected with a Gal4-driven luciferase reporter as shown in the schematic with increasing amounts of Gal4DBD-ORC1 or Gal4DBD-SUV39H1 together with pCMV-LacZ plasmids. Relative luciferase activity was determined and normalized to lacZ activity. Experiments were carried out in triplicate. The whole cell extract was immunoblotted with anti-Gal4 antibody for expression of Gal4DBD fusion plasmids. α-Tubulin served as a loading control. Statistical analysis was performed using the Student’s t test. **p

    Article Snippet: Plasmid construction and mutagenesis Plasmids expressing GFP-RB, 10–4 CCNE1 (Addgene: Cyclin E gene) promoter, E2F1, DP1, HDAC1 and pGL2-GAL4-UAS-Luc were purchased from Addgene. pCMV-LacZ plasmid was purchased from Clontech.

    Techniques: Transfection, Luciferase, Activity Assay, Expressing

    The DPE and MTE are important regulators of transcription from the slob71 promoter. Core nucleotides within the MTE, DPE, or both MTE and DPE were mutated in the slob71 −1966 to +81 promoter. Luciferase activity was measured in Drosophila S2 cells transfected with mutant or intact control constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the percentage of activity exhibited by the intact control construct. *** p

    Journal: The Journal of Neuroscience

    Article Title: Cell-Specific Fine-Tuning of Neuronal Excitability by Differential Expression of Modulator Protein Isoforms

    doi: 10.1523/JNEUROSCI.1001-13.2013

    Figure Lengend Snippet: The DPE and MTE are important regulators of transcription from the slob71 promoter. Core nucleotides within the MTE, DPE, or both MTE and DPE were mutated in the slob71 −1966 to +81 promoter. Luciferase activity was measured in Drosophila S2 cells transfected with mutant or intact control constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the percentage of activity exhibited by the intact control construct. *** p

    Article Snippet: S2 cells were transfected with the luciferase reporter constructs (2 μg/well) along with a vector expressing β-galactosidase (β-gal; pCMV–LacZ vector from Clontech; 1 μg/well) in duplicate using Lipofectamine (Invitrogen).

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

    Promoter elements in specific domains of slob71 affect transcriptional activity. A , Promoter fragments of slob71 were inserted upstream of a minP in the pGL4.23[ luc2 /minP] vector, which drives a low level of basal luciferase expression. Luciferase activity was measured in Drosophila S2 cells transfected with minP–luc or slob71 promoter fragment–minP–luc constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the fold change compared with the empty minP–luc vector. B , C , Core nucleotides within the HB and MIRR recognition sites were mutated in slob71 promoters. Luciferase activity was measured in Drosophila S2 cells transfected with mutant or intact control constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the percentage of activity exhibited by the intact control construct. Mutating MIRR or HB sites in the slob71 −1966 to +81 promoter increases relative luciferase activity ( B ). Mutation of the HB site in the slob71 −1966 to −1500 promoter fragment upstream of the minP increases relative luciferase activity, whereas mutation of the MIRR site has no effect ( C ). *** p

    Journal: The Journal of Neuroscience

    Article Title: Cell-Specific Fine-Tuning of Neuronal Excitability by Differential Expression of Modulator Protein Isoforms

    doi: 10.1523/JNEUROSCI.1001-13.2013

    Figure Lengend Snippet: Promoter elements in specific domains of slob71 affect transcriptional activity. A , Promoter fragments of slob71 were inserted upstream of a minP in the pGL4.23[ luc2 /minP] vector, which drives a low level of basal luciferase expression. Luciferase activity was measured in Drosophila S2 cells transfected with minP–luc or slob71 promoter fragment–minP–luc constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the fold change compared with the empty minP–luc vector. B , C , Core nucleotides within the HB and MIRR recognition sites were mutated in slob71 promoters. Luciferase activity was measured in Drosophila S2 cells transfected with mutant or intact control constructs. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the percentage of activity exhibited by the intact control construct. Mutating MIRR or HB sites in the slob71 −1966 to +81 promoter increases relative luciferase activity ( B ). Mutation of the HB site in the slob71 −1966 to −1500 promoter fragment upstream of the minP increases relative luciferase activity, whereas mutation of the MIRR site has no effect ( C ). *** p

    Article Snippet: S2 cells were transfected with the luciferase reporter constructs (2 μg/well) along with a vector expressing β-galactosidase (β-gal; pCMV–LacZ vector from Clontech; 1 μg/well) in duplicate using Lipofectamine (Invitrogen).

    Techniques: Activity Assay, Plasmid Preparation, Luciferase, Expressing, Transfection, Construct, Mutagenesis

    slob57 and slob71 promoters exhibit different transcriptional activity. Promoter regions upstream of the identified TSSs for slob57 and slob71 were cloned into the pGL4.10[ luc2 ] vector to drive the luciferase (luc) reporter gene. Drosophila S2 cells were transfected with various slob promoter–luc constructs and the pCMV–LacZ vector as an internal control. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the fold change compared with the empty luc control vector. A , Summary of luciferase activity experiments with slob57 promoters. B , Summary of luciferase activity experiments with slob71 promoters. Relative luciferase activity driven by slob71 promoters is higher than that of slob57 . * p

    Journal: The Journal of Neuroscience

    Article Title: Cell-Specific Fine-Tuning of Neuronal Excitability by Differential Expression of Modulator Protein Isoforms

    doi: 10.1523/JNEUROSCI.1001-13.2013

    Figure Lengend Snippet: slob57 and slob71 promoters exhibit different transcriptional activity. Promoter regions upstream of the identified TSSs for slob57 and slob71 were cloned into the pGL4.10[ luc2 ] vector to drive the luciferase (luc) reporter gene. Drosophila S2 cells were transfected with various slob promoter–luc constructs and the pCMV–LacZ vector as an internal control. The relative luciferase activity is the luciferase activity normalized to β-gal activity and is reported as the fold change compared with the empty luc control vector. A , Summary of luciferase activity experiments with slob57 promoters. B , Summary of luciferase activity experiments with slob71 promoters. Relative luciferase activity driven by slob71 promoters is higher than that of slob57 . * p

    Article Snippet: S2 cells were transfected with the luciferase reporter constructs (2 μg/well) along with a vector expressing β-galactosidase (β-gal; pCMV–LacZ vector from Clontech; 1 μg/well) in duplicate using Lipofectamine (Invitrogen).

    Techniques: Activity Assay, Clone Assay, Plasmid Preparation, Luciferase, Transfection, Construct

    Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with pCMV backbone, pCMV lacZ, enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment

    doi: 10.1038/mtm.2014.12

    Figure Lengend Snippet: Antitumor effects of nontherapeutic plasmid. ( a ) Tumor growth curve of CT26 tumors treated with pCMV backbone, pCMV lacZ, enhanced expression vector (pEEV) backbone, pEEV lacZ, and an untreated tumor growing on Balb/c mice ( n = 6 per group) is presented as tumor diameter measured. Tumor volume was calculated as previously described. Data presented are means ± SEM of six individual tumors. *Compared to the untreated group and pEEV lacZ, * P

    Article Snippet: Plasmids For plasmid transfection, a pCMV-lacz plasmid (Plasmid factory) was used encoding a β-galactosidase protein (LacZ) gene under the control of a CMV promoter.

    Techniques: Plasmid Preparation, Expressing, Mouse Assay

    β-galactosidase expression in murine tissue. ( a ) Copy numbers of lacZ transgene ascertained by quantitative polymerase chain reaction (PCR) in murine tissue. Bar graph showing absolute copy number of the lacz transgene per nanogram of genomic DNA 2 days after electroporation obtained from MF1-nu/nu mouse tissue. All gDNA samples were normalized to 100 ng of DNA prior to PCR. Each individual sample was analyzed in triplicate for each quantiative PCR (qPCR). qPCR values are means ± standard error of the mean (SEM) of triplicate measurements. A comparison of enhanced expression vector (pEEV) (light gray bars) and pCMV (dark gray bars) samples were performed. *** P

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment

    doi: 10.1038/mtm.2014.12

    Figure Lengend Snippet: β-galactosidase expression in murine tissue. ( a ) Copy numbers of lacZ transgene ascertained by quantitative polymerase chain reaction (PCR) in murine tissue. Bar graph showing absolute copy number of the lacz transgene per nanogram of genomic DNA 2 days after electroporation obtained from MF1-nu/nu mouse tissue. All gDNA samples were normalized to 100 ng of DNA prior to PCR. Each individual sample was analyzed in triplicate for each quantiative PCR (qPCR). qPCR values are means ± standard error of the mean (SEM) of triplicate measurements. A comparison of enhanced expression vector (pEEV) (light gray bars) and pCMV (dark gray bars) samples were performed. *** P

    Article Snippet: Plasmids For plasmid transfection, a pCMV-lacz plasmid (Plasmid factory) was used encoding a β-galactosidase protein (LacZ) gene under the control of a CMV promoter.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Electroporation, Plasmid Preparation

    β-Galactosidase expression in porcine tissue. ( a ) Representative image of β-galactosidase staining in porcine tissues. β-Galactosidase expression significantly increased in enhanced expression vector (pEEV) lacZ in all tissues been examined. (b) Evaluation of lacZ mRNA transcript expression by qRT-PCR in porcine tissue. Bar graph presenting the relative expression of the lacZ mRNA 2 days after electroporation. All qPCR data were normalized using 18S RNA as reference gene. Relative expression levels are plotted as means ± standard error of the mean (SEM) of triplicate measurements. pEEV (light gray bars) and pCMV (dark gray bars). pEEV lacZ expressed was significantly higher than pCMV lacZ. ( c ) Copy numbers of lacZ transgene ascertained by quantitative PCR in porcine tissue. Bar graph showing absolute copy number of the lacz transgene per nanogram of genomic DNAs 2 days after electroporation. All gDNA samples were normalized to 100 ng of DNA prior to PCR. Each individual sample was analyzed in triplicate for each qPCR. qPCR values are means ± SEM of triplicate measurements. A comparison of pEEV (light gray bars) and pCMV (dark gray bars) samples were performed.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment

    doi: 10.1038/mtm.2014.12

    Figure Lengend Snippet: β-Galactosidase expression in porcine tissue. ( a ) Representative image of β-galactosidase staining in porcine tissues. β-Galactosidase expression significantly increased in enhanced expression vector (pEEV) lacZ in all tissues been examined. (b) Evaluation of lacZ mRNA transcript expression by qRT-PCR in porcine tissue. Bar graph presenting the relative expression of the lacZ mRNA 2 days after electroporation. All qPCR data were normalized using 18S RNA as reference gene. Relative expression levels are plotted as means ± standard error of the mean (SEM) of triplicate measurements. pEEV (light gray bars) and pCMV (dark gray bars). pEEV lacZ expressed was significantly higher than pCMV lacZ. ( c ) Copy numbers of lacZ transgene ascertained by quantitative PCR in porcine tissue. Bar graph showing absolute copy number of the lacz transgene per nanogram of genomic DNAs 2 days after electroporation. All gDNA samples were normalized to 100 ng of DNA prior to PCR. Each individual sample was analyzed in triplicate for each qPCR. qPCR values are means ± SEM of triplicate measurements. A comparison of pEEV (light gray bars) and pCMV (dark gray bars) samples were performed.

    Article Snippet: Plasmids For plasmid transfection, a pCMV-lacz plasmid (Plasmid factory) was used encoding a β-galactosidase protein (LacZ) gene under the control of a CMV promoter.

    Techniques: Expressing, Staining, Plasmid Preparation, Quantitative RT-PCR, Electroporation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Growth curve for E. coli DH5α-pCMV-lacZ . The growth curve based on OD 600 measurements is marked by small black circles and the appropriate best-fit curve is indicated in blue. The harvesting time points T 1 , T 2 , and T 3 are presented as bold big points. The minimum–maximum area, within which the measurement values have to reside, is bordered by the red curve (minimal measured OD 600 value) and the green curve (maximal measured OD 600 value).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation

    doi: 10.3389/fbioe.2015.00127

    Figure Lengend Snippet: Growth curve for E. coli DH5α-pCMV-lacZ . The growth curve based on OD 600 measurements is marked by small black circles and the appropriate best-fit curve is indicated in blue. The harvesting time points T 1 , T 2 , and T 3 are presented as bold big points. The minimum–maximum area, within which the measurement values have to reside, is bordered by the red curve (minimal measured OD 600 value) and the green curve (maximal measured OD 600 value).

    Article Snippet: Bacterial strain and plasmids The Escherichia coli strain DH5α (F–Φ80lac ZΔM15 Δ(lac ZYA-arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

    Techniques:

    HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from pCMV-lacZ. Each bar on the graph represents the average of four separate transfections.

    Journal: Zebrafish

    Article Title: A Zebrafish Cell Culture Assay for the Identification of MicroRNA Targets

    doi: 10.1089/zeb.2010.0674

    Figure Lengend Snippet: HEK293 cells fail to support the interaction of zebrafish miR-430b with known mRNA targets. (A) Time course of miR-430b-mediated inhibition of luciferase expression using the reporter construct pGL3-3xPT, which carries three tandom copies of the perfect target sequence. pGL3-3xPT and miR-430b were cotransfected into the HEK293 cells and luciferase expression was measured at the times indicated post-transfection. (B) miR-430b-mediated inhibition of luciferase expression using reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, and pGl3-3′nanos that carry 3′UTRs containing known miR-430b target sequences. Luciferase expression was measured at 48 h post-transfection (hpt). pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively, and the transfection efficiency was normalized to β-galactosidase expression from pCMV-lacZ. Each bar on the graph represents the average of four separate transfections.

    Article Snippet: On the second day the medium was changed and transfection complex consisting of the Lipofectamine2000 reagent (2 μL), miRNA (50 nM), pCMV-lacZ (250 ng; Clontech), pGL3 plasmid (500 ng), and OPTI-MEMI (280 μL total volume; Gibco) was added to each well.

    Techniques: Inhibition, Luciferase, Expressing, Construct, Sequencing, Transfection

    ZSSJ cells support the interaction of zebrafish miR-430b and mRNA target sequences. ZSSJ cells were transfected with miR-430b or control miRNA along with one of three luciferase reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, or pGl3-3′nanos, that carry known 3′UTR target sequences for miR-430b. Luciferase expression was measured 48 hpt. pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively. The transfection efficiency was normalized to β-galactosidase expression from pCMV-lacZ. Each bar represents the average of three separate transfections. *A significant difference ( p

    Journal: Zebrafish

    Article Title: A Zebrafish Cell Culture Assay for the Identification of MicroRNA Targets

    doi: 10.1089/zeb.2010.0674

    Figure Lengend Snippet: ZSSJ cells support the interaction of zebrafish miR-430b and mRNA target sequences. ZSSJ cells were transfected with miR-430b or control miRNA along with one of three luciferase reporter constructs, pGl3-3′zgc63829, pGl3-3′gstm, or pGl3-3′nanos, that carry known 3′UTR target sequences for miR-430b. Luciferase expression was measured 48 hpt. pGL3-3xPT and pGL-3 promoter were used as positive and negative controls, respectively. The transfection efficiency was normalized to β-galactosidase expression from pCMV-lacZ. Each bar represents the average of three separate transfections. *A significant difference ( p

    Article Snippet: On the second day the medium was changed and transfection complex consisting of the Lipofectamine2000 reagent (2 μL), miRNA (50 nM), pCMV-lacZ (250 ng; Clontech), pGL3 plasmid (500 ng), and OPTI-MEMI (280 μL total volume; Gibco) was added to each well.

    Techniques: Transfection, Luciferase, Construct, Expressing

    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

    Fusion assay between ASCT2 -transduced and syncytin-Ory1 -transduced co-cultured cells demonstrates that ASCT2 is the syncytin-Ory1 receptor . Left panel: Cell-cell fusion was assayed upon independent transfections of a set of A23 cells with an empty vector (none) or an expression vector for either the syncytin-Ory1, syncytin-1 or syncytin-2 protein together with an nls- LacZ gene-expression vector, and another set of A23 cells with an expression vector for the syncytin-1 receptor ASCT2, the syncytin-2 receptor MFSD2 [ 13 ] or an empty vector (none). One day after transfection, cells were resuspended and pairs of transfected cells from each set were cocultured for 1-2 days, fixed and X-Gal stained. Right panel: Syncytia can be easily detected (arrows) for the syncytin-Ory1/ASCT2, syncytin-1/ASCT2 and syncytin-2/MFSD2 pairs, with only mononucleated cells visible in the other cases. Abbreviations: syn-Ory1, syncytin-Ory1; syn1, syncytin-1; syn2, syncytin-2.

    Journal: Retrovirology

    Article Title: Identification of an endogenous retroviral envelope gene with fusogenic activity and placenta-specific expression in the rabbit: a new "syncytin" in a third order of mammals

    doi: 10.1186/1742-4690-6-107

    Figure Lengend Snippet: Fusion assay between ASCT2 -transduced and syncytin-Ory1 -transduced co-cultured cells demonstrates that ASCT2 is the syncytin-Ory1 receptor . Left panel: Cell-cell fusion was assayed upon independent transfections of a set of A23 cells with an empty vector (none) or an expression vector for either the syncytin-Ory1, syncytin-1 or syncytin-2 protein together with an nls- LacZ gene-expression vector, and another set of A23 cells with an expression vector for the syncytin-1 receptor ASCT2, the syncytin-2 receptor MFSD2 [ 13 ] or an empty vector (none). One day after transfection, cells were resuspended and pairs of transfected cells from each set were cocultured for 1-2 days, fixed and X-Gal stained. Right panel: Syncytia can be easily detected (arrows) for the syncytin-Ory1/ASCT2, syncytin-1/ASCT2 and syncytin-2/MFSD2 pairs, with only mononucleated cells visible in the other cases. Abbreviations: syn-Ory1, syncytin-Ory1; syn1, syncytin-1; syn2, syncytin-2.

    Article Snippet: For the self-fusion assay, cells seeded at 104 - 5 × 104 cells per well in 24-well plates were transfected by using the Lipofectamine kit (Invitrogen) with 0.2 μg of either the syncytin-Ory1 expressing or an empty vector, supplemented with 0.2 μg of a LacZ-expression vector (pCMV-β, Clontech).

    Techniques: Single Vesicle Fusion Assay, Cell Culture, Transfection, Plasmid Preparation, Expressing, Staining

    Fusogenic activity of syncytin-Ory1 . (A) Assay for cell-cell fusion mediated by syncytin-Ory1. The indicated cell lines were transfected with an expression vector for syncytin-Ory1 or an empty vector (none) together with a LacZ expression vector. Cells were cultured for 1-2 days after transfection, fixed and X-gal-stained. Syncytia (arrows) were detected in syncytin-Ory1 -transfected SH-SY5Y cells, with only mononucleated cells visible in the other cases. (B) Assay for cell infection mediated by syncytin-Ory1-pseudotyped virus particles. Pseudotypes were produced by cotransfection of human 293T cells with expression vectors for the SIV core, the syncytin-Ory1 protein (or an empty vector) and a LacZ -containing retroviral transcript. Supernatants were used to infect the indicated target cells, which were X-gal stained 3 days after infection. Abbreviation: Syn-Ory1, syncytin-Ory1.

    Journal: Retrovirology

    Article Title: Identification of an endogenous retroviral envelope gene with fusogenic activity and placenta-specific expression in the rabbit: a new "syncytin" in a third order of mammals

    doi: 10.1186/1742-4690-6-107

    Figure Lengend Snippet: Fusogenic activity of syncytin-Ory1 . (A) Assay for cell-cell fusion mediated by syncytin-Ory1. The indicated cell lines were transfected with an expression vector for syncytin-Ory1 or an empty vector (none) together with a LacZ expression vector. Cells were cultured for 1-2 days after transfection, fixed and X-gal-stained. Syncytia (arrows) were detected in syncytin-Ory1 -transfected SH-SY5Y cells, with only mononucleated cells visible in the other cases. (B) Assay for cell infection mediated by syncytin-Ory1-pseudotyped virus particles. Pseudotypes were produced by cotransfection of human 293T cells with expression vectors for the SIV core, the syncytin-Ory1 protein (or an empty vector) and a LacZ -containing retroviral transcript. Supernatants were used to infect the indicated target cells, which were X-gal stained 3 days after infection. Abbreviation: Syn-Ory1, syncytin-Ory1.

    Article Snippet: For the self-fusion assay, cells seeded at 104 - 5 × 104 cells per well in 24-well plates were transfected by using the Lipofectamine kit (Invitrogen) with 0.2 μg of either the syncytin-Ory1 expressing or an empty vector, supplemented with 0.2 μg of a LacZ-expression vector (pCMV-β, Clontech).

    Techniques: Activity Assay, Transfection, Expressing, Plasmid Preparation, Cell Culture, Staining, Infection, Produced, Cotransfection