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  • 98
    New England Biolabs agei
    Agei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 902 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher ecori
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    TaKaRa ecori
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    nhei  (TaKaRa)
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    TaKaRa nhei
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    TaKaRa bglii
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    TaKaRa pcr amplification
    Pcr Amplification, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 21524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa in fusion hd cloning kit
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    TaKaRa pmcherry n1
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    xhoi  (TaKaRa)
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    TaKaRa xhoi
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    kpni  (TaKaRa)
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    TaKaRa kpni
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    TaKaRa hindiii
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    TaKaRa pmcherry c1
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    TaKaRa puromycin resistance gene
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    Thermo Fisher lipofectamine 2000
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    mlui  (TaKaRa)
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    TaKaRa mlui
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    TaKaRa pdsred2 er
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    TaKaRa xbai
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    TaKaRa 293t cells
    Design and development of NINJA targeting construct. a, Schematic of NINJA targeting construct (NINJA TC) after integration into Rosa26 (R26) locus. NINJA contains regulatory (RM) and neoantigen (NM) modules. Schematic shows modules after Cre or FLPo recombinase and doxycycline/tamoxifen (D/T) exposure. (Red and white fill arrow pairs are non-compatible loxP sites. Black and white ball and stick lines are splice sites, and black or grey arrows indicate promoters) b-c, FLPo exposure is required for GFP expression from NM. <t>293T</t> cells were transfected with the indicated constructs and assessed by FACS or western blotting. b, Histogram shows GFP expression 72 hours after transfection with NM.7 alone (gray, filled) or NM.7 and FLPo (line). Percent GFP+ for each is indicated. c, Western blotting for GRP94 (control, top panel) or N-terminal GFP (bottom panel) on lysates from 293T cells. Positive control (+) is cell lysate from KP-C4A3D6 after FLPo (see Fig. 2 ). d-e, NM is only immunogenic after FLPo exposure. DC2.4 cells were transfected with NM.7 or NM.7+FLPo and cultured with or without naive Cell Trace Violet (CTV)-stained CD8 P14 T cells. d, Tumor cell wells were stained with crystal violet four days later. e, FACS plots show CTV dilution and CD44 expression on CD8+ Va2+ T cells. f, NINJA TC expresses GFP after Cre, rtTA, and D/T treatment. 293T cells were transfected with NINJA TC with and without plasmids expressing Cre and rtTA. Cells were then cultured with or without D/T. Histograms show GFP expression from the 293T cells with the indicated conditions.
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    ndei  (TaKaRa)
    99
    TaKaRa ndei
    Design and development of NINJA targeting construct. a, Schematic of NINJA targeting construct (NINJA TC) after integration into Rosa26 (R26) locus. NINJA contains regulatory (RM) and neoantigen (NM) modules. Schematic shows modules after Cre or FLPo recombinase and doxycycline/tamoxifen (D/T) exposure. (Red and white fill arrow pairs are non-compatible loxP sites. Black and white ball and stick lines are splice sites, and black or grey arrows indicate promoters) b-c, FLPo exposure is required for GFP expression from NM. <t>293T</t> cells were transfected with the indicated constructs and assessed by FACS or western blotting. b, Histogram shows GFP expression 72 hours after transfection with NM.7 alone (gray, filled) or NM.7 and FLPo (line). Percent GFP+ for each is indicated. c, Western blotting for GRP94 (control, top panel) or N-terminal GFP (bottom panel) on lysates from 293T cells. Positive control (+) is cell lysate from KP-C4A3D6 after FLPo (see Fig. 2 ). d-e, NM is only immunogenic after FLPo exposure. DC2.4 cells were transfected with NM.7 or NM.7+FLPo and cultured with or without naive Cell Trace Violet (CTV)-stained CD8 P14 T cells. d, Tumor cell wells were stained with crystal violet four days later. e, FACS plots show CTV dilution and CD44 expression on CD8+ Va2+ T cells. f, NINJA TC expresses GFP after Cre, rtTA, and D/T treatment. 293T cells were transfected with NINJA TC with and without plasmids expressing Cre and rtTA. Cells were then cultured with or without D/T. Histograms show GFP expression from the 293T cells with the indicated conditions.
    Ndei, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa pdsred2 c1
    Design and development of NINJA targeting construct. a, Schematic of NINJA targeting construct (NINJA TC) after integration into Rosa26 (R26) locus. NINJA contains regulatory (RM) and neoantigen (NM) modules. Schematic shows modules after Cre or FLPo recombinase and doxycycline/tamoxifen (D/T) exposure. (Red and white fill arrow pairs are non-compatible loxP sites. Black and white ball and stick lines are splice sites, and black or grey arrows indicate promoters) b-c, FLPo exposure is required for GFP expression from NM. <t>293T</t> cells were transfected with the indicated constructs and assessed by FACS or western blotting. b, Histogram shows GFP expression 72 hours after transfection with NM.7 alone (gray, filled) or NM.7 and FLPo (line). Percent GFP+ for each is indicated. c, Western blotting for GRP94 (control, top panel) or N-terminal GFP (bottom panel) on lysates from 293T cells. Positive control (+) is cell lysate from KP-C4A3D6 after FLPo (see Fig. 2 ). d-e, NM is only immunogenic after FLPo exposure. DC2.4 cells were transfected with NM.7 or NM.7+FLPo and cultured with or without naive Cell Trace Violet (CTV)-stained CD8 P14 T cells. d, Tumor cell wells were stained with crystal violet four days later. e, FACS plots show CTV dilution and CD44 expression on CD8+ Va2+ T cells. f, NINJA TC expresses GFP after Cre, rtTA, and D/T treatment. 293T cells were transfected with NINJA TC with and without plasmids expressing Cre and rtTA. Cells were then cultured with or without D/T. Histograms show GFP expression from the 293T cells with the indicated conditions.
    Pdsred2 C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 424 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Design and development of NINJA targeting construct. a, Schematic of NINJA targeting construct (NINJA TC) after integration into Rosa26 (R26) locus. NINJA contains regulatory (RM) and neoantigen (NM) modules. Schematic shows modules after Cre or FLPo recombinase and doxycycline/tamoxifen (D/T) exposure. (Red and white fill arrow pairs are non-compatible loxP sites. Black and white ball and stick lines are splice sites, and black or grey arrows indicate promoters) b-c, FLPo exposure is required for GFP expression from NM. 293T cells were transfected with the indicated constructs and assessed by FACS or western blotting. b, Histogram shows GFP expression 72 hours after transfection with NM.7 alone (gray, filled) or NM.7 and FLPo (line). Percent GFP+ for each is indicated. c, Western blotting for GRP94 (control, top panel) or N-terminal GFP (bottom panel) on lysates from 293T cells. Positive control (+) is cell lysate from KP-C4A3D6 after FLPo (see Fig. 2 ). d-e, NM is only immunogenic after FLPo exposure. DC2.4 cells were transfected with NM.7 or NM.7+FLPo and cultured with or without naive Cell Trace Violet (CTV)-stained CD8 P14 T cells. d, Tumor cell wells were stained with crystal violet four days later. e, FACS plots show CTV dilution and CD44 expression on CD8+ Va2+ T cells. f, NINJA TC expresses GFP after Cre, rtTA, and D/T treatment. 293T cells were transfected with NINJA TC with and without plasmids expressing Cre and rtTA. Cells were then cultured with or without D/T. Histograms show GFP expression from the 293T cells with the indicated conditions.

    Journal: bioRxiv

    Article Title: NINJA: an inducible genetic model for creating neoantigens in vivo

    doi: 10.1101/2020.01.09.900894

    Figure Lengend Snippet: Design and development of NINJA targeting construct. a, Schematic of NINJA targeting construct (NINJA TC) after integration into Rosa26 (R26) locus. NINJA contains regulatory (RM) and neoantigen (NM) modules. Schematic shows modules after Cre or FLPo recombinase and doxycycline/tamoxifen (D/T) exposure. (Red and white fill arrow pairs are non-compatible loxP sites. Black and white ball and stick lines are splice sites, and black or grey arrows indicate promoters) b-c, FLPo exposure is required for GFP expression from NM. 293T cells were transfected with the indicated constructs and assessed by FACS or western blotting. b, Histogram shows GFP expression 72 hours after transfection with NM.7 alone (gray, filled) or NM.7 and FLPo (line). Percent GFP+ for each is indicated. c, Western blotting for GRP94 (control, top panel) or N-terminal GFP (bottom panel) on lysates from 293T cells. Positive control (+) is cell lysate from KP-C4A3D6 after FLPo (see Fig. 2 ). d-e, NM is only immunogenic after FLPo exposure. DC2.4 cells were transfected with NM.7 or NM.7+FLPo and cultured with or without naive Cell Trace Violet (CTV)-stained CD8 P14 T cells. d, Tumor cell wells were stained with crystal violet four days later. e, FACS plots show CTV dilution and CD44 expression on CD8+ Va2+ T cells. f, NINJA TC expresses GFP after Cre, rtTA, and D/T treatment. 293T cells were transfected with NINJA TC with and without plasmids expressing Cre and rtTA. Cells were then cultured with or without D/T. Histograms show GFP expression from the 293T cells with the indicated conditions.

    Article Snippet: We also confirmed that the correctly spliced products were generated by generating cDNA from 293T cells transfected with the NM.1 construct with and without FLPo. cDNA was amplified using using PrimeSTAR HS DNA Polymerase (Takara Bio Inc., cat. R040A) and the primers GFP F AgeI 72 and GP80 R XhoI 64 ( ).

    Techniques: Construct, Expressing, Transfection, FACS, Western Blot, Positive Control, Cell Culture, Staining