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  • 95
    Addgene inc ps pax2
    Induction of the TRIM5α-activated antiviral state does not require a productive infection. (a) Representative FACS dot plots of TRIM5-KO or control Jurkat cells infected or not with AT-2- or MetOH (vehicle)-treated NRC10 DsRed , followed by challenge with NRC1 GFP 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control Jurkat cells with NRC1 GFP or NRC10 GFP, 48 h after a first infection with NRC10 DsRed or EC8-2 DsRed treated with AT-2 or with the vehicle only (M). Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. (b) Representative dot plots of TRIM5-KO and control THP-1 cells infected or not with AT-2- or vehicle-treated NRC10 DsRed , or with “empty” <t>psPAX2-vector</t> containing NRC10 CA (NRC10_EV), followed by NRC1 GFP challenge 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control THP-1 cells with NRC1 GFP, 48 h after a first infection using NRC10 DsRed or EC5 DsRed treated with AT-2, with the vehicle only, or using NRC10 or EC5-2 “empty vectors”. Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. Shown in all bar graphs of this figure are representative experiments performed in triplicates and repeated at least three times. Individual values are included (black symbols).
    Ps Pax2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 573 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Addgene inc packaging plasmid ps pax2 2
    Validation of the integration-defective phenotype of vector particles generated with the aid of packaging construct <t>psPAX2.IN</t> D116N . (A) Genetic composition of the HIV-1-based LVs and IDLVs generated for and used in the current study. Lentiviral vectors
    Packaging Plasmid Ps Pax2 2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 87/100, based on 253 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Addgene inc pspax 2 lentiviral helper plasmids
    Validation of the integration-defective phenotype of vector particles generated with the aid of packaging construct <t>psPAX2.IN</t> D116N . (A) Genetic composition of the HIV-1-based LVs and IDLVs generated for and used in the current study. Lentiviral vectors
    Pspax 2 Lentiviral Helper Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Addgene inc plasmids pspax 2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Plasmids Pspax 2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Addgene inc generation packaging plasmids pspax 2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Generation Packaging Plasmids Pspax 2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Crown Bioscience pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2, supplied by Crown Bioscience, used in various techniques. Bioz Stars score: 95/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cellecta pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2, supplied by Cellecta, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 206 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Addgene inc lentiviral pspax 2 packaging
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Lentiviral Pspax 2 Packaging, supplied by Addgene inc, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Addgene inc pspax 2 packaging plasmid
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax 2 Packaging Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Addgene inc pspax2 d64v
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2 D64v, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Addgene inc lenticrispr pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Lenticrispr Pspax2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Addgene inc generation 2 0 lentiviral packaging plasmids pspax 2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Generation 2 0 Lentiviral Packaging Plasmids Pspax 2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc pspax2 vector
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2 Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Addgene inc plnb vectors pspax2
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Plnb Vectors Pspax2, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc pspax2 packing plasmids
    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs <t>psPAX2</t> VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).
    Pspax2 Packing Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Induction of the TRIM5α-activated antiviral state does not require a productive infection. (a) Representative FACS dot plots of TRIM5-KO or control Jurkat cells infected or not with AT-2- or MetOH (vehicle)-treated NRC10 DsRed , followed by challenge with NRC1 GFP 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control Jurkat cells with NRC1 GFP or NRC10 GFP, 48 h after a first infection with NRC10 DsRed or EC8-2 DsRed treated with AT-2 or with the vehicle only (M). Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. (b) Representative dot plots of TRIM5-KO and control THP-1 cells infected or not with AT-2- or vehicle-treated NRC10 DsRed , or with “empty” psPAX2-vector containing NRC10 CA (NRC10_EV), followed by NRC1 GFP challenge 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control THP-1 cells with NRC1 GFP, 48 h after a first infection using NRC10 DsRed or EC5 DsRed treated with AT-2, with the vehicle only, or using NRC10 or EC5-2 “empty vectors”. Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. Shown in all bar graphs of this figure are representative experiments performed in triplicates and repeated at least three times. Individual values are included (black symbols).

    Journal: PLoS Pathogens

    Article Title: HIV-1 capsids from B27/B57+ elite controllers escape Mx2 but are targeted by TRIM5α, leading to the induction of an antiviral state

    doi: 10.1371/journal.ppat.1007398

    Figure Lengend Snippet: Induction of the TRIM5α-activated antiviral state does not require a productive infection. (a) Representative FACS dot plots of TRIM5-KO or control Jurkat cells infected or not with AT-2- or MetOH (vehicle)-treated NRC10 DsRed , followed by challenge with NRC1 GFP 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control Jurkat cells with NRC1 GFP or NRC10 GFP, 48 h after a first infection with NRC10 DsRed or EC8-2 DsRed treated with AT-2 or with the vehicle only (M). Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. (b) Representative dot plots of TRIM5-KO and control THP-1 cells infected or not with AT-2- or vehicle-treated NRC10 DsRed , or with “empty” psPAX2-vector containing NRC10 CA (NRC10_EV), followed by NRC1 GFP challenge 48 h later. The % of GFP + cells are shown in bold. Bar graphs showing the percentage of infected cells following infection of T5KO or control THP-1 cells with NRC1 GFP, 48 h after a first infection using NRC10 DsRed or EC5 DsRed treated with AT-2, with the vehicle only, or using NRC10 or EC5-2 “empty vectors”. Means with SD are plotted. Numbers on the bars represent the -fold inhibition relative to the relevant mock control. Shown in all bar graphs of this figure are representative experiments performed in triplicates and repeated at least three times. Individual values are included (black symbols).

    Article Snippet: Capsid sequences were amplified from NL4-3, NRC10, EC5-2, EC8-2 and EC9-2 using FOR_PsPAX2_ClaI and REV_PsPAX2_ECORV were ligated into psPAX2 (Addgene #12260) cut with EcoRV and ClaI.

    Techniques: Infection, FACS, Inhibition, Plasmid Preparation

    Indirect immunofluorescence microscopy demonstrates that LDLR expression induces rerouting of VSVG rather than progression to the Golgi apparatus. HEK293 cells were transfected with psPAX2 and pMD2.G with or without RC200006 and probed for VSVG and GM130 (A). Note colocalization in the absence of LDLR (yellow). (B) Cells probed for VSVG and ERGIC53. Note colocalization in the presence of LDLR. (C) Cells probed for VSVG and LAMP2. Note the VSVG-granules enclosed by LAMP2-positive membranes in the presence of LDLR. (D) Cells probed for VSVG and vimentin. Note the VSVG-granules surrounded by cages of collapsed vimentin in the presence of LDLR. In any case, prominent surface expression of VSVG was only seen in the absence of LDLR expression. The micrographs in panels A, B, and C were taken using a Zeiss Axioplan 2 microscope at 100× magnification and deconvolved using the deconvolution filter of the G'MIC plugin package for GIMP software. The micrographs in panel D were acquired by using a confocal microscope at 100× magnification. Scale bars, 10 μm.

    Journal: Journal of Virology

    Article Title: Release of Vesicular Stomatitis Virus Spike Protein G-Pseudotyped Lentivirus from the Host Cell Is Impaired upon Low-Density Lipoprotein Receptor Overexpression

    doi: 10.1128/JVI.01869-15

    Figure Lengend Snippet: Indirect immunofluorescence microscopy demonstrates that LDLR expression induces rerouting of VSVG rather than progression to the Golgi apparatus. HEK293 cells were transfected with psPAX2 and pMD2.G with or without RC200006 and probed for VSVG and GM130 (A). Note colocalization in the absence of LDLR (yellow). (B) Cells probed for VSVG and ERGIC53. Note colocalization in the presence of LDLR. (C) Cells probed for VSVG and LAMP2. Note the VSVG-granules enclosed by LAMP2-positive membranes in the presence of LDLR. (D) Cells probed for VSVG and vimentin. Note the VSVG-granules surrounded by cages of collapsed vimentin in the presence of LDLR. In any case, prominent surface expression of VSVG was only seen in the absence of LDLR expression. The micrographs in panels A, B, and C were taken using a Zeiss Axioplan 2 microscope at 100× magnification and deconvolved using the deconvolution filter of the G'MIC plugin package for GIMP software. The micrographs in panel D were acquired by using a confocal microscope at 100× magnification. Scale bars, 10 μm.

    Article Snippet: To confirm the above observation and determine the underlying reason, virus was produced in HEK293 cells (2.5 × 106 /10-cm2 plate) via transient transfection with the self-inactivating vector construct pHR-CMV-EGFP (3.33 μg; Addgene 14858) , as well as the packaging construct psPAX2 (5 μg; Addgene 12260) and the envelope construct pMD2.G, encoding the VSVG surface protein (1.66 μg; Addgene 12259) (both a kind gift from Didier Trono).

    Techniques: Immunofluorescence, Microscopy, Expressing, Transfection, Software

    Lentiviral copy number increases with decreasing amount of LDLR-encoding plasmid cotransfected with the virus-producing plasmids used in the positive control. (A) Viral RNA in the supernatant of HEK293 cells was determined by RT-qPCR. The negative controls (NegC) were psPAX2 and pMD2.G, and the positive controls (PosC) were pHR-CMV-EGFP, psPAX2, and pMD2.G. Data are the arithmetic means of 3 biological replicates ± standard errors of the means (SEM). The presence of LDLR decreases the copy number significantly [one-way analysis of variance (ANOVA), F (4, 2) = 356.18], and the copy number is inversely related to the concentration of LDLR [one-way ANOVA, F(3, 2) = 373.33]. Wedges indicate the amount of the LDLR-encoding plasmid relative to pHR-CMV-EGFP used in the transfection (LDLR, 1:2 to 1:16). ***, P

    Journal: Journal of Virology

    Article Title: Release of Vesicular Stomatitis Virus Spike Protein G-Pseudotyped Lentivirus from the Host Cell Is Impaired upon Low-Density Lipoprotein Receptor Overexpression

    doi: 10.1128/JVI.01869-15

    Figure Lengend Snippet: Lentiviral copy number increases with decreasing amount of LDLR-encoding plasmid cotransfected with the virus-producing plasmids used in the positive control. (A) Viral RNA in the supernatant of HEK293 cells was determined by RT-qPCR. The negative controls (NegC) were psPAX2 and pMD2.G, and the positive controls (PosC) were pHR-CMV-EGFP, psPAX2, and pMD2.G. Data are the arithmetic means of 3 biological replicates ± standard errors of the means (SEM). The presence of LDLR decreases the copy number significantly [one-way analysis of variance (ANOVA), F (4, 2) = 356.18], and the copy number is inversely related to the concentration of LDLR [one-way ANOVA, F(3, 2) = 373.33]. Wedges indicate the amount of the LDLR-encoding plasmid relative to pHR-CMV-EGFP used in the transfection (LDLR, 1:2 to 1:16). ***, P

    Article Snippet: To confirm the above observation and determine the underlying reason, virus was produced in HEK293 cells (2.5 × 106 /10-cm2 plate) via transient transfection with the self-inactivating vector construct pHR-CMV-EGFP (3.33 μg; Addgene 14858) , as well as the packaging construct psPAX2 (5 μg; Addgene 12260) and the envelope construct pMD2.G, encoding the VSVG surface protein (1.66 μg; Addgene 12259) (both a kind gift from Didier Trono).

    Techniques: Plasmid Preparation, Positive Control, Quantitative RT-PCR, Concentration Assay, Transfection

    Validation of the integration-defective phenotype of vector particles generated with the aid of packaging construct psPAX2.IN D116N . (A) Genetic composition of the HIV-1-based LVs and IDLVs generated for and used in the current study. Lentiviral vectors

    Journal: Human Gene Therapy

    Article Title: Histone Deacetylase Inhibition Activates Transgene Expression from Integration-Defective Lentiviral Vectors in Dividing and Non-Dividing Cells

    doi: 10.1089/hum.2012.069

    Figure Lengend Snippet: Validation of the integration-defective phenotype of vector particles generated with the aid of packaging construct psPAX2.IN D116N . (A) Genetic composition of the HIV-1-based LVs and IDLVs generated for and used in the current study. Lentiviral vectors

    Article Snippet: The second-generation packaging plasmid psPAX2 was provided by Didier Trono (Addgene plasmid 12260; Cambridge, MA), whilst the third-generation packaging plasmid pLP1, the HIV-1 rev expression plasmid pLP2, and the vesicular stomatitis glycoprotein-G (VSV-G) pseudotyping construct pLP/VSVG are from Invitrogen.

    Techniques: Plasmid Preparation, Generated, Construct

    Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs psPAX2 VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).

    Journal: Journal of Virology

    Article Title: Sensing of HIV-1 Entry Triggers a Type I Interferon Response in Human Primary Macrophages

    doi: 10.1128/JVI.00147-17

    Figure Lengend Snippet: Fusion is required to trigger macrophage response to HIV-1. (A) MDM were infected with HIV-1 ΔEnv VSVG or HIV-1 NL-AD8 at 1 μg/ml in the presence of AZT or the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 6). (B) MDM were infected with HIV-1 132W (maintained and produced in MT4C5) in the presence or the absence of the fusion inhibitor T20. Cells were assayed at 24 hpi for intracellular Nef and viperin mRNA expression (3 independent experiments, n = 9). (C) MDM were infected with HIV-1 ΔEnv VSVG or VLPs psPAX2 VSVG , corresponding to equivalents of 0.1 μg/ml and 1 μg/ml of p24. Cells were assayed at 24 hpi for viperin mRNA. Entry of virus and VLP was confirmed by intracellular Gag staining. FACS plots from one representative donor are shown, and data were compiled from 3 independent experiments ( n = 6). (D) MDM were infected with HIV-1 ΔEnv VSVG , HIV-1 ΔEnv ADA , or VLPs psPAX2 ADA at 0.1 μg/ml and 1 μg/ml for 24 h. Viral entry was confirmed by intracellular Gag staining (2 independent experiments, n = 4).

    Article Snippet: HIV-1 NL4-3ΔEnv (kindly provided by O. Schwartz, Institut Pasteur, Paris, France), HIV-1 NL-AD8 , and psPAX2 VLP were produced by transfection of the corresponding proviral DNA or psPAX2 plasmid (12260; Addgene) in HEK 293T cells (ATCC number CRL-11268) by polyethyleneimine (PEI) precipitation.

    Techniques: Infection, Expressing, Produced, Staining, FACS