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  • 99
    Thermo Fisher packaging plasmids
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Packaging Plasmids, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3178 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 3178 article reviews
    Price from $9.99 to $1999.99
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    99
    Addgene inc packaging plasmids pspax2
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Packaging Plasmids Pspax2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 1138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids pspax2/product/Addgene inc
    Average 99 stars, based on 1138 article reviews
    Price from $9.99 to $1999.99
    packaging plasmids pspax2 - by Bioz Stars, 2020-11
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    93
    Addgene inc packaging plasmids
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Packaging Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids/product/Addgene inc
    Average 93 stars, based on 1524 article reviews
    Price from $9.99 to $1999.99
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    94
    Addgene inc pspax2 packaging plasmid
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Pspax2 Packaging Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 485 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pspax2 packaging plasmid/product/Addgene inc
    Average 94 stars, based on 485 article reviews
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    99
    Addgene inc packaging plasmids pmd2 g
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Packaging Plasmids Pmd2 G, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 696 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    System Biosciences Inc ppack packaging plasmid mix
    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon <t>lentiviral-mediated</t> AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.
    Ppack Packaging Plasmid Mix, supplied by System Biosciences Inc, used in various techniques. Bioz Stars score: 89/100, based on 392 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ppack packaging plasmid mix/product/System Biosciences Inc
    Average 89 stars, based on 392 article reviews
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    91
    Addgene inc lentiviral packaging plasmids
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Lentiviral Packaging Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 181 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Addgene inc packaging plasmids pcmv dr8 2 dvpr
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Packaging Plasmids Pcmv Dr8 2 Dvpr, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids pcmv dr8 2 dvpr/product/Addgene inc
    Average 99 stars, based on 37 article reviews
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    99
    Addgene inc packaging plasmids pcmv vsv g
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Packaging Plasmids Pcmv Vsv G, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids pcmv vsv g/product/Addgene inc
    Average 99 stars, based on 192 article reviews
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    88
    Stratagene pvpack vsv g packaging plasmids
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Pvpack Vsv G Packaging Plasmids, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvpack vsv g packaging plasmids/product/Stratagene
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    88
    System Biosciences Inc ppackh1 packaging plasmid mix
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Ppackh1 Packaging Plasmid Mix, supplied by System Biosciences Inc, used in various techniques. Bioz Stars score: 88/100, based on 143 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ppackh1 packaging plasmid mix/product/System Biosciences Inc
    Average 88 stars, based on 143 article reviews
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    99
    Addgene inc packaging plasmids pmdlg prre
    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with <t>lentiviral</t> shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P
    Packaging Plasmids Pmdlg Prre, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids pmdlg prre/product/Addgene inc
    Average 99 stars, based on 80 article reviews
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    90
    Addgene inc lentivirus packaging plasmids
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Lentivirus Packaging Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Addgene inc packaging plasmids pcmv dr8 2
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmids Pcmv Dr8 2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 88/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Mirus Bio packaging plasmids
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmids, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 93/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 248 article reviews
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    99
    Addgene inc packaging plasmids prsv rev
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmids Prsv Rev, supplied by Addgene inc, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids prsv rev/product/Addgene inc
    Average 99 stars, based on 51 article reviews
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    93
    OriGene packaging plasmids
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmids, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 203 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmids/product/OriGene
    Average 93 stars, based on 203 article reviews
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    packaging plasmids - by Bioz Stars, 2020-11
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    91
    Addgene inc packaging plasmid pcmv vsvg
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmid Pcmv Vsvg, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/packaging plasmid pcmv vsvg/product/Addgene inc
    Average 91 stars, based on 32 article reviews
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    92
    Genecopoeia packaging plasmids
    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding <t>lentivirus</t> formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.
    Packaging Plasmids, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 92/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 88 article reviews
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    Image Search Results


    AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon lentiviral-mediated AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.

    Journal: Journal of Lipid Research

    Article Title: Acid ceramidase promotes drug resistance in acute myeloid leukemia through NF-κB-dependent P-glycoprotein upregulation

    doi: 10.1194/jlr.M091876

    Figure Lengend Snippet: AC knockdown decreased P-gp protein expression. A: Western blot showing the reduction of AC and P-gp levels upon lentiviral-mediated AC knockdown (72 h) in drug-resistant HL-60/VCR, HL-60/ABTR, KG1/ABTR, and KG1a/ABTR cell lines compared with GFP control vector. B: AC and P-gp bands were quantified and normalized to β-actin. The blots and graph are representative of two independent experiments.

    Article Snippet: Plasmids were transfected into HEK293 T/17 cells with lentiviral packaging plasmids (Invitrogen).

    Techniques: Expressing, Western Blot, Plasmid Preparation

    Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with lentiviral shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P

    Journal: Nucleic Acids Research

    Article Title: Dual roles of c-Myc in the regulation of hTERT gene

    doi: 10.1093/nar/gku721

    Figure Lengend Snippet: Effect of c-Myc overexpression on hTERT promoter induction. ( A ) c-Myc KD in Tel + /H(wt) cells containing LNCX-c-Myc (Myc) or LNCX vector (Neo) retroviruses. The cells were transduced with lentiviral shRNAs and harvested for western analysis 4 days later. ( B ) Induction of transgenic hTERT promoter. Luciferase activities were measured 4 days post infection and hTERT promoter activities were determined as Rluc / Fluc . **, P

    Article Snippet: Control pLKO.1 plasmids, vector and scrambled shRNAs and lentiviral packaging plasmids were obtained from Addgene (Cambridge, MA).

    Techniques: Over Expression, Plasmid Preparation, Transduction, Western Blot, Transgenic Assay, Luciferase, Infection

    Effects of EBP knockdown on hTERT regulation. ( A ) Knockdown of EBPs by shRNAs. Tel + cells were infected with lentiviral shRNAs against EBPs for 4 days. Protein expression was determined by western analyses using antibodies as indicated. Upper panels are EBPs and lower panels show actin as a loading control. ( B ) Regulation of chromatinized hTERT reporter by shRNAs. Tel + H(wt) cells were infected with lentiviral shRNA and luciferase activities were measured 2, 3 and 4 days post infection. hTERT promoter activities were determined as Rluc / Fluc . Vec, lentiviral vector pLKO.1; SCR, scrambled shRNA; Unt, no infection.

    Journal: Nucleic Acids Research

    Article Title: Dual roles of c-Myc in the regulation of hTERT gene

    doi: 10.1093/nar/gku721

    Figure Lengend Snippet: Effects of EBP knockdown on hTERT regulation. ( A ) Knockdown of EBPs by shRNAs. Tel + cells were infected with lentiviral shRNAs against EBPs for 4 days. Protein expression was determined by western analyses using antibodies as indicated. Upper panels are EBPs and lower panels show actin as a loading control. ( B ) Regulation of chromatinized hTERT reporter by shRNAs. Tel + H(wt) cells were infected with lentiviral shRNA and luciferase activities were measured 2, 3 and 4 days post infection. hTERT promoter activities were determined as Rluc / Fluc . Vec, lentiviral vector pLKO.1; SCR, scrambled shRNA; Unt, no infection.

    Article Snippet: Control pLKO.1 plasmids, vector and scrambled shRNAs and lentiviral packaging plasmids were obtained from Addgene (Cambridge, MA).

    Techniques: Infection, Expressing, Western Blot, shRNA, Luciferase, Plasmid Preparation

    Effects of c-Myc KD on TF recruitment to the hTERT promoters. Tel + H(wt) cells were infected with lentiviral shRNAs against c-Myc, c-MycA9 and c-MycA10, for 4 days. ( A ) The expression of TFs. mRNA levels were measured by quantitative RT-PCR and normalized to 18S rRNA. ( B ) Binding of TFs to the hTERT promoters. ChIP experiments were performed as described in Figure 6 . *, P

    Journal: Nucleic Acids Research

    Article Title: Dual roles of c-Myc in the regulation of hTERT gene

    doi: 10.1093/nar/gku721

    Figure Lengend Snippet: Effects of c-Myc KD on TF recruitment to the hTERT promoters. Tel + H(wt) cells were infected with lentiviral shRNAs against c-Myc, c-MycA9 and c-MycA10, for 4 days. ( A ) The expression of TFs. mRNA levels were measured by quantitative RT-PCR and normalized to 18S rRNA. ( B ) Binding of TFs to the hTERT promoters. ChIP experiments were performed as described in Figure 6 . *, P

    Article Snippet: Control pLKO.1 plasmids, vector and scrambled shRNAs and lentiviral packaging plasmids were obtained from Addgene (Cambridge, MA).

    Techniques: Infection, Expressing, Quantitative RT-PCR, Binding Assay, Chromatin Immunoprecipitation

    Effects on mutant hTERT promoters by EBP knockdown. Tel + H(EboxD) and Tel + H(EboxDM) cells were infected with lentiviral shRNAs. Luciferase activities were measured 2, 3 and 4 days after infection. hTERT promoter activities were determined as Rluc / Fluc .

    Journal: Nucleic Acids Research

    Article Title: Dual roles of c-Myc in the regulation of hTERT gene

    doi: 10.1093/nar/gku721

    Figure Lengend Snippet: Effects on mutant hTERT promoters by EBP knockdown. Tel + H(EboxD) and Tel + H(EboxDM) cells were infected with lentiviral shRNAs. Luciferase activities were measured 2, 3 and 4 days after infection. hTERT promoter activities were determined as Rluc / Fluc .

    Article Snippet: Control pLKO.1 plasmids, vector and scrambled shRNAs and lentiviral packaging plasmids were obtained from Addgene (Cambridge, MA).

    Techniques: Mutagenesis, Infection, Luciferase

    Effect of c-Myc/Max knockdown on hTERT expression in human epithelial cell line MCF10A. MCF10A cells were infected with lentiviral shRNAs and selected with 1 μg/ml puromycin for 4 days. hTERT mRNA levels were determined by quantitative RT-PCR and normalized to 18S rRNA. *, p

    Journal: Nucleic Acids Research

    Article Title: Dual roles of c-Myc in the regulation of hTERT gene

    doi: 10.1093/nar/gku721

    Figure Lengend Snippet: Effect of c-Myc/Max knockdown on hTERT expression in human epithelial cell line MCF10A. MCF10A cells were infected with lentiviral shRNAs and selected with 1 μg/ml puromycin for 4 days. hTERT mRNA levels were determined by quantitative RT-PCR and normalized to 18S rRNA. *, p

    Article Snippet: Control pLKO.1 plasmids, vector and scrambled shRNAs and lentiviral packaging plasmids were obtained from Addgene (Cambridge, MA).

    Techniques: Expressing, Infection, Quantitative RT-PCR

    Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding lentivirus formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.

    Journal: Cell Death & Disease

    Article Title: A long non-coding RNA interacts with Gfra1 and maintains survival of mouse spermatogonial stem cells

    doi: 10.1038/cddis.2016.24

    Figure Lengend Snippet: Reduced repopulation capacity of SSCs after lncRNA033862 knockdown. ( a and b ) SSCs transduced with shRNA1894-encoding lentivirus formed significantly fewer spermatogenic colonies in recipient testes than SSCs transduced with vector control, scale bar =1mm. Data are from three independent transplantation experiments; in total, control vector lentivirus transduced SSC were transplanted into 21 testes, and sh1894 lentivirus transduced SSC into 22 testes. Donor-derived colonies exhibited a full range of germ cell development, including mature spermatids (black arrows, scale bar=1 mm for left panel and 20 μ m for right panel), irrespective of lncRNA033862 knockdown. ( c ) Proposed model for the regulation of mouse SSC self-renewal and maintenance by lncRNA033862. Under normal physiologic conditions in the testes, exogenous growth factor GDNF binds to its receptor GFRA1 and RET and stimulates intracellular signaling involving PI3K/AKT and ETK pathways. This positively regulates transcription of lncRNA033862, such that sufficient transcript levels are present in the nucleus to activate the transcription of genes necessary for SSC self-renewal, such as Gfra1 . ( d ) In the absence of GDNF, lower lncRNA033862 transcription, leading to reduced activation of Gfra1 transcription, causes imbalance of self-renewal and maintenance of SSCs. Error bars indicate S.D.

    Article Snippet: Lentivirus particles were generated by co-transfection of shRNA plasmids and lentivirus packaging plasmids into HEK293T cells using CaCl2 transfection following a protocol provided by Addgene ( http://www.addgene.org/tools/protocols/plko/ ).

    Techniques: Transduction, Plasmid Preparation, Transplantation Assay, Derivative Assay, Activation Assay

    Knockdown of LncRNA 033862 impairs the survival and proliferation of SSCs . (a) In vitro lentivirus-mediated knockdown resulted in a reduction of lncRNA033862 transcript levels by 30% with shRNA663, and by 65% by shRNA1894, and a corresponding reduction of Gfra1 transcript levels ( b ) to 88.9 % with shRNA663, and to 40.9% with shRNA1894. ( c ) Significant reduction of total cell number in SSC cultures 7 days after transduction with shRNA663 and shRNA1894 compared with vector alone (shVector). FACS analysis of SSCs gated according to forward and side scatter as indicated in ( d ). ( e and f ) Compared with shVector transfection, knockdown lncRNA033862 by sh1894 resulted in an increase of apoptotic cells in SSC cultures. ( g ) Lentiviral transduction of SSCs with shRNA663 or shRNA1894 resulted in an increased number of dead cells and ( h ) significantly higher total numbers of apoptotic cells in SSCs transduced with shRNA1894. ( i ) Both shRNA663 ( P

    Journal: Cell Death & Disease

    Article Title: A long non-coding RNA interacts with Gfra1 and maintains survival of mouse spermatogonial stem cells

    doi: 10.1038/cddis.2016.24

    Figure Lengend Snippet: Knockdown of LncRNA 033862 impairs the survival and proliferation of SSCs . (a) In vitro lentivirus-mediated knockdown resulted in a reduction of lncRNA033862 transcript levels by 30% with shRNA663, and by 65% by shRNA1894, and a corresponding reduction of Gfra1 transcript levels ( b ) to 88.9 % with shRNA663, and to 40.9% with shRNA1894. ( c ) Significant reduction of total cell number in SSC cultures 7 days after transduction with shRNA663 and shRNA1894 compared with vector alone (shVector). FACS analysis of SSCs gated according to forward and side scatter as indicated in ( d ). ( e and f ) Compared with shVector transfection, knockdown lncRNA033862 by sh1894 resulted in an increase of apoptotic cells in SSC cultures. ( g ) Lentiviral transduction of SSCs with shRNA663 or shRNA1894 resulted in an increased number of dead cells and ( h ) significantly higher total numbers of apoptotic cells in SSCs transduced with shRNA1894. ( i ) Both shRNA663 ( P

    Article Snippet: Lentivirus particles were generated by co-transfection of shRNA plasmids and lentivirus packaging plasmids into HEK293T cells using CaCl2 transfection following a protocol provided by Addgene ( http://www.addgene.org/tools/protocols/plko/ ).

    Techniques: In Vitro, Transduction, Plasmid Preparation, FACS, Transfection