retroviral transduction Search Results


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  • 99
    ATCC retroviral transduction
    Live cell imaging-based analysis of NK cell cytotoxicity. (A) Representative time-lapse images of interaction between <t>NK-92-CD16</t> cells (yellow lines) and HN-31 cells (white lines) treated with various antibodies. (B–D) Effects of antibody treatment on overall cytotoxicity (B), time for killing (C), and number of cancer cells killed by an NK cell (D). Mann-Whitney test was used. *P
    Retroviral Transduction, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher retroviral transduction 293ft cells
    Live cell imaging-based analysis of NK cell cytotoxicity. (A) Representative time-lapse images of interaction between <t>NK-92-CD16</t> cells (yellow lines) and HN-31 cells (white lines) treated with various antibodies. (B–D) Effects of antibody treatment on overall cytotoxicity (B), time for killing (C), and number of cancer cells killed by an NK cell (D). Mann-Whitney test was used. *P
    Retroviral Transduction 293ft Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc retroviral transduction
    Live cell imaging-based analysis of NK cell cytotoxicity. (A) Representative time-lapse images of interaction between <t>NK-92-CD16</t> cells (yellow lines) and HN-31 cells (white lines) treated with various antibodies. (B–D) Effects of antibody treatment on overall cytotoxicity (B), time for killing (C), and number of cancer cells killed by an NK cell (D). Mann-Whitney test was used. *P
    Retroviral Transduction, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson retroviral transduction
    Efficient transfer of the DMF5 TCR to peripheral blood lymphocyes and in vitro efficacy of DMF5 T cells. (A) MART-1 pentamer staining of T cells transduced with the <t>retroviral</t> vector encoding the DMF5 recombinant TCR. (B) IFN-γ release by DMF5 T cells and mock T cells after coculturing with HLA-A*02 + melanoma cell line Mel624 and HLA-A*02 − Mel888, and plate-bound anti-CD3/anti-CD28 as a control. (C) IFN-γ release by DMF5 T cells and mock T cells after coculturing with peptide-pulsed T2 cells and plate-bound anti-CD3/anti-CD28 as a control. (D) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with peptide-pulsed T2 cells. (E) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with HLA-A*02 + melanoma cell line Mel624, and HLA-A*02 − melanoma cell line Mel888. TCR, T cell receptors.
    Retroviral Transduction, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 338 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc retrovirus transduction
    Efficient transfer of the DMF5 TCR to peripheral blood lymphocyes and in vitro efficacy of DMF5 T cells. (A) MART-1 pentamer staining of T cells transduced with the <t>retroviral</t> vector encoding the DMF5 recombinant TCR. (B) IFN-γ release by DMF5 T cells and mock T cells after coculturing with HLA-A*02 + melanoma cell line Mel624 and HLA-A*02 − Mel888, and plate-bound anti-CD3/anti-CD28 as a control. (C) IFN-γ release by DMF5 T cells and mock T cells after coculturing with peptide-pulsed T2 cells and plate-bound anti-CD3/anti-CD28 as a control. (D) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with peptide-pulsed T2 cells. (E) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with HLA-A*02 + melanoma cell line Mel624, and HLA-A*02 − melanoma cell line Mel888. TCR, T cell receptors.
    Retrovirus Transduction, supplied by Addgene inc, used in various techniques. Bioz Stars score: 89/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Addgene inc retroviral transduction vector pbabepuro
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Retroviral Transduction Vector Pbabepuro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa retroviral transduction
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Retroviral Transduction, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 699 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Biolabs Inc transduction pmxs mir gfp puro retroviral expression vector
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Transduction Pmxs Mir Gfp Puro Retroviral Expression Vector, supplied by Cell Biolabs Inc, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc transduction retroviral gfp vector
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Transduction Retroviral Gfp Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc retrovirus transduction pmig mgata3
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Retrovirus Transduction Pmig Mgata3, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC retrovirus transduction 293t cells
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Retrovirus Transduction 293t Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Addgene inc retrovirus transduction plncx akt vector
    <t>Retroviral</t> transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with <t>pBABEpuro–p53C132F/E168G</t> or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.
    Retrovirus Transduction Plncx Akt Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc retrovirus transduction plncx akt vectors
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Retrovirus Transduction Plncx Akt Vectors, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC retrovirus transduced nih3t3 cells
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Retrovirus Transduced Nih3t3 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Orbigen phoenixtm retroviral transduction system
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Phoenixtm Retroviral Transduction System, supplied by Orbigen, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    TaKaRa plxsn retroviral vector
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Plxsn Retroviral Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Addgene inc pmxs retroviral transduction
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Pmxs Retroviral Transduction, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa retro x q retroviral vector transduction system
    <t>AKT</t> is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty <t>retrovirus,</t> or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.
    Retro X Q Retroviral Vector Transduction System, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC retroviral transduction mda mb 231 breast carcinoma
    Cell spreading and directed migration is impaired in CD9/CD81-depleted <t>MDA-MB-231.</t> ( A ) MDA-MB-231 Wild type, CD9/CD81si, CD9RX, and CD81RX cells were plated on LM-332-coated glass coverslips, and cell spreading was imaged 30 min later by phase contrast microscopy. ( B ) The area of cell spreading for each cell type was calculated by subtracting the mean area of cells imaged immediately after plating from the mean area of cells after 30 min of spreading. Values are means ± s.e.m.; n = 3 trials with at least 25 cells of each cell type per trial; *P
    Retroviral Transduction Mda Mb 231 Breast Carcinoma, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc retroviral transduction over expression
    Autophagic flux in virus-specific CD8 T cells is inversely correlated with cell proliferation status. ( a ) MSCV <t>retroviral</t> constructs showing the transgenes (GFP-LC3b and GFP-LC3b-G120A) introduced in P14 cells to probe for autophagy activity. ( b ) Flow cytometry plots of adoptively transferred P14 cells in spleens. P14 cells transduced with retroviral plasmid of either MIT-GFP-LC3b or MIT-G120A are positive for the congenic marker Thy1.1. Day 5, 8 and 30 p.i. splenocytes were used for the analysis. The percentage of GFP-negative cells out of the transduced P14 (Thy1.1 + ) cells from each group is highlighted in blue on the lower right corner of each plot and is summarized in ( c ). ( d ) Representative ImageStream ® data showing images of P14 cells from either GFP-LC3b or G120A groups on day 8 p.i.. Images showing GFP signals of GFP + MIT vector-transduced P14 cells (Ly5.1 + Thy1.1 + ). Summary graph of ImageStream ® analysis is plotted in ( e ) showing percentage of GFP + or Thy1.1 + P14 cells that exhibited more than one GFP punctum. Errors bars represent SEM. ( b ) and ( d ) are representative of three independent experiments, n≥3 mice in each group.
    Retroviral Transduction Over Expression, supplied by Addgene inc, used in various techniques. Bioz Stars score: 88/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher retroviral transduction lipofectamine 2000
    Autophagic flux in virus-specific CD8 T cells is inversely correlated with cell proliferation status. ( a ) MSCV <t>retroviral</t> constructs showing the transgenes (GFP-LC3b and GFP-LC3b-G120A) introduced in P14 cells to probe for autophagy activity. ( b ) Flow cytometry plots of adoptively transferred P14 cells in spleens. P14 cells transduced with retroviral plasmid of either MIT-GFP-LC3b or MIT-G120A are positive for the congenic marker Thy1.1. Day 5, 8 and 30 p.i. splenocytes were used for the analysis. The percentage of GFP-negative cells out of the transduced P14 (Thy1.1 + ) cells from each group is highlighted in blue on the lower right corner of each plot and is summarized in ( c ). ( d ) Representative ImageStream ® data showing images of P14 cells from either GFP-LC3b or G120A groups on day 8 p.i.. Images showing GFP signals of GFP + MIT vector-transduced P14 cells (Ly5.1 + Thy1.1 + ). Summary graph of ImageStream ® analysis is plotted in ( e ) showing percentage of GFP + or Thy1.1 + P14 cells that exhibited more than one GFP punctum. Errors bars represent SEM. ( b ) and ( d ) are representative of three independent experiments, n≥3 mice in each group.
    Retroviral Transduction Lipofectamine 2000, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC retroviral transduction human saos 2
    Autophagic flux in virus-specific CD8 T cells is inversely correlated with cell proliferation status. ( a ) MSCV <t>retroviral</t> constructs showing the transgenes (GFP-LC3b and GFP-LC3b-G120A) introduced in P14 cells to probe for autophagy activity. ( b ) Flow cytometry plots of adoptively transferred P14 cells in spleens. P14 cells transduced with retroviral plasmid of either MIT-GFP-LC3b or MIT-G120A are positive for the congenic marker Thy1.1. Day 5, 8 and 30 p.i. splenocytes were used for the analysis. The percentage of GFP-negative cells out of the transduced P14 (Thy1.1 + ) cells from each group is highlighted in blue on the lower right corner of each plot and is summarized in ( c ). ( d ) Representative ImageStream ® data showing images of P14 cells from either GFP-LC3b or G120A groups on day 8 p.i.. Images showing GFP signals of GFP + MIT vector-transduced P14 cells (Ly5.1 + Thy1.1 + ). Summary graph of ImageStream ® analysis is plotted in ( e ) showing percentage of GFP + or Thy1.1 + P14 cells that exhibited more than one GFP punctum. Errors bars represent SEM. ( b ) and ( d ) are representative of three independent experiments, n≥3 mice in each group.
    Retroviral Transduction Human Saos 2, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Live cell imaging-based analysis of NK cell cytotoxicity. (A) Representative time-lapse images of interaction between NK-92-CD16 cells (yellow lines) and HN-31 cells (white lines) treated with various antibodies. (B–D) Effects of antibody treatment on overall cytotoxicity (B), time for killing (C), and number of cancer cells killed by an NK cell (D). Mann-Whitney test was used. *P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Anti-tumor effects of NK cells and anti-PD-L1 antibody with antibody-dependent cellular cytotoxicity in PD-L1-positive cancer cell lines

    doi: 10.1136/jitc-2020-000873

    Figure Lengend Snippet: Live cell imaging-based analysis of NK cell cytotoxicity. (A) Representative time-lapse images of interaction between NK-92-CD16 cells (yellow lines) and HN-31 cells (white lines) treated with various antibodies. (B–D) Effects of antibody treatment on overall cytotoxicity (B), time for killing (C), and number of cancer cells killed by an NK cell (D). Mann-Whitney test was used. *P

    Article Snippet: NK-92-CD16 cells (purchased from the ATCC, PTA-8836) were maintained in alpha minimum essential medium supplemented with 25% FBS, 0.2 mM inositol, 0.1 mM 2­mercaptoethanol, and 0.02 mM folic acid.

    Techniques: Live Cell Imaging, MANN-WHITNEY

    Anti-PD-L1 mAbs-mediated ADCC in human cancer cell lines. NK-92-CD16 cytotoxicity against tumor cells was measured by a standard 51 Cr-release assay with various E:T ratios (30:1, 10:1, and 3:1). Bar graph represents cytotoxicity of NK-92-CD16 cells against cancer cell lines at 30:1 E:T ratio. All cancer cell lines were treated with 10 µg/mL of IgG1 isotype control (black dotted lines and bar), atezolizumab (green), IMC-001 (red), and anti-hPD-L1-hIgG1 (blue). (A) Head and neck squamous cell carcinoma cell lines. PD-L1 low (left) includes SNU-1041, SNU-1066, and Detroit 562, and PD-L1 high (right) includes SNU-1076, FaDu, and HN31 cells. (B) For non-small-cell lung cancer, NCI-H1650 cells are PD-L1 low (left), and NCI-H1975 cells are PD-L1 high (right). All experiments were performed three times independently. Statistical significance across groups 6 was determined by one-way analysis of variance. All data are shown as mean±SD. *P

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Anti-tumor effects of NK cells and anti-PD-L1 antibody with antibody-dependent cellular cytotoxicity in PD-L1-positive cancer cell lines

    doi: 10.1136/jitc-2020-000873

    Figure Lengend Snippet: Anti-PD-L1 mAbs-mediated ADCC in human cancer cell lines. NK-92-CD16 cytotoxicity against tumor cells was measured by a standard 51 Cr-release assay with various E:T ratios (30:1, 10:1, and 3:1). Bar graph represents cytotoxicity of NK-92-CD16 cells against cancer cell lines at 30:1 E:T ratio. All cancer cell lines were treated with 10 µg/mL of IgG1 isotype control (black dotted lines and bar), atezolizumab (green), IMC-001 (red), and anti-hPD-L1-hIgG1 (blue). (A) Head and neck squamous cell carcinoma cell lines. PD-L1 low (left) includes SNU-1041, SNU-1066, and Detroit 562, and PD-L1 high (right) includes SNU-1076, FaDu, and HN31 cells. (B) For non-small-cell lung cancer, NCI-H1650 cells are PD-L1 low (left), and NCI-H1975 cells are PD-L1 high (right). All experiments were performed three times independently. Statistical significance across groups 6 was determined by one-way analysis of variance. All data are shown as mean±SD. *P

    Article Snippet: NK-92-CD16 cells (purchased from the ATCC, PTA-8836) were maintained in alpha minimum essential medium supplemented with 25% FBS, 0.2 mM inositol, 0.1 mM 2­mercaptoethanol, and 0.02 mM folic acid.

    Techniques: Release Assay

    Efficient transfer of the DMF5 TCR to peripheral blood lymphocyes and in vitro efficacy of DMF5 T cells. (A) MART-1 pentamer staining of T cells transduced with the retroviral vector encoding the DMF5 recombinant TCR. (B) IFN-γ release by DMF5 T cells and mock T cells after coculturing with HLA-A*02 + melanoma cell line Mel624 and HLA-A*02 − Mel888, and plate-bound anti-CD3/anti-CD28 as a control. (C) IFN-γ release by DMF5 T cells and mock T cells after coculturing with peptide-pulsed T2 cells and plate-bound anti-CD3/anti-CD28 as a control. (D) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with peptide-pulsed T2 cells. (E) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with HLA-A*02 + melanoma cell line Mel624, and HLA-A*02 − melanoma cell line Mel888. TCR, T cell receptors.

    Journal: Human Gene Therapy Methods

    Article Title: Potential Limitations of the NSG Humanized Mouse as a Model System to Optimize Engineered Human T cell Therapy for Cancer

    doi: 10.1089/hgtb.2013.022

    Figure Lengend Snippet: Efficient transfer of the DMF5 TCR to peripheral blood lymphocyes and in vitro efficacy of DMF5 T cells. (A) MART-1 pentamer staining of T cells transduced with the retroviral vector encoding the DMF5 recombinant TCR. (B) IFN-γ release by DMF5 T cells and mock T cells after coculturing with HLA-A*02 + melanoma cell line Mel624 and HLA-A*02 − Mel888, and plate-bound anti-CD3/anti-CD28 as a control. (C) IFN-γ release by DMF5 T cells and mock T cells after coculturing with peptide-pulsed T2 cells and plate-bound anti-CD3/anti-CD28 as a control. (D) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with peptide-pulsed T2 cells. (E) Cytotoxicity by CD107a expression of DMF5 T cells when cocultured with HLA-A*02 + melanoma cell line Mel624, and HLA-A*02 − melanoma cell line Mel888. TCR, T cell receptors.

    Article Snippet: For retroviral transduction, non-tissue-culture six-well plates (BD Biosciences) were coated with retronectin (10 μg/ml; Takara, Invitrogen) and incubated at 4°C overnight.

    Techniques: In Vitro, Staining, Transduction, Plasmid Preparation, Recombinant, Expressing

    Effects of IL-7 and IL-15 on the proliferation, function, and phenotype of DMF5 T cells. (A) Proliferation of T cells cultured in IL-2, IL-7, and IL-15 after being activated with anti-CD3 and anti-CD28 antibodies, and transduced with the retroviral vector encoding the DMF5 TCR. (B) Viability of DMF5 T cells cultured in IL-2, IL-7, and IL-15 measured by flow cytometry according to the percentage of cells within the live lymphocyte gate. (C–F) Expression of costimulatory molecules CD27 and CD28 (C and D) , and markers of homing to central lymphoid organs CCR7 and CD62L (E and F) on CD8 + DMF5 T cells after being cultured in IL-2, IL-7, and IL-15. (G) Representative flow cytometry plots showing production of IFN-γ and IL-2 by CD8 + DMF5 T cells cultured in IL-2, IL-7, or IL-15 and stimulated with T2 cells loaded with MART-1 peptide. (H) Cytotoxicity of DMF5 T cells cultured in IL-2, IL-7, or IL-15 by CD107a expression after coculture with HLA-A*02 + cell lines Mel624, Mel501, and WM2664, and HLA-A*02 − cell line Mel888. p -Values

    Journal: Human Gene Therapy Methods

    Article Title: Potential Limitations of the NSG Humanized Mouse as a Model System to Optimize Engineered Human T cell Therapy for Cancer

    doi: 10.1089/hgtb.2013.022

    Figure Lengend Snippet: Effects of IL-7 and IL-15 on the proliferation, function, and phenotype of DMF5 T cells. (A) Proliferation of T cells cultured in IL-2, IL-7, and IL-15 after being activated with anti-CD3 and anti-CD28 antibodies, and transduced with the retroviral vector encoding the DMF5 TCR. (B) Viability of DMF5 T cells cultured in IL-2, IL-7, and IL-15 measured by flow cytometry according to the percentage of cells within the live lymphocyte gate. (C–F) Expression of costimulatory molecules CD27 and CD28 (C and D) , and markers of homing to central lymphoid organs CCR7 and CD62L (E and F) on CD8 + DMF5 T cells after being cultured in IL-2, IL-7, and IL-15. (G) Representative flow cytometry plots showing production of IFN-γ and IL-2 by CD8 + DMF5 T cells cultured in IL-2, IL-7, or IL-15 and stimulated with T2 cells loaded with MART-1 peptide. (H) Cytotoxicity of DMF5 T cells cultured in IL-2, IL-7, or IL-15 by CD107a expression after coculture with HLA-A*02 + cell lines Mel624, Mel501, and WM2664, and HLA-A*02 − cell line Mel888. p -Values

    Article Snippet: For retroviral transduction, non-tissue-culture six-well plates (BD Biosciences) were coated with retronectin (10 μg/ml; Takara, Invitrogen) and incubated at 4°C overnight.

    Techniques: Cell Culture, Transduction, Plasmid Preparation, Flow Cytometry, Cytometry, Expressing

    Retroviral transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with pBABEpuro–p53C132F/E168G or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.

    Journal: Frontiers in Oncology

    Article Title: Adaptive Resistance to Immunotherapy Directed Against p53 Can be Overcome by Global Expression of Tumor-Antigens in Dendritic Cells

    doi: 10.3389/fonc.2014.00270

    Figure Lengend Snippet: Retroviral transduction of D2SC/1 cells . (A) D2SC/1 cells were cultured in the supernatant of the retroviral packaging cell line FLYA4lacZ3 for 24 h and LacZ transduced cells were visualized by X-gal staining. (B) The mp53 retroviral transduction vectors are illustrated. Predicted K[d] restricted peptide presentation is marked by a box. Amino acids at position 2, 3, 5, and carboxyl termini function as anchors or auxiliary anchors within the MHC pocket. (C) Flow cytometry of mp53-transduced D2SC/1. Stable amphotropic packaging cell lines were generated by calcium phosphate transfection of FLY-AF-13 cells with pBABEpuro–p53C132F/E168G or pBABEpuro–p53M234I and puromycin selection before D2SC/1 cells were transduced three times with 1.5–3 × 10 4 CFU/ml of budding virus for 15 h. Cells were fixed and stained with 1 μg/ml PAb 421 anti-p53 antibody. Filled histograms represent stained cells transduced with the pBABEpuro control vector, open histograms D2SC/1-p53C132F/E168G, or D2SC/1-p53M234I transduced cells.

    Article Snippet: The missense point mutations C132F, E168G, and M234I present in the p53 alleles were verified by sequence analysis. p53M234I and p53C132F/E168G were cloned into the retroviral transduction vector pBABEpuro (Addgene, Cambridge, MA, USA).

    Techniques: Transduction, Cell Culture, Staining, Flow Cytometry, Cytometry, Generated, Transfection, Selection, Plasmid Preparation

    AKT is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty retrovirus, or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.

    Journal: Oncotarget

    Article Title: Human papillomavirus type 18 E5 oncogene supports cell cycle progression and impairs epithelial differentiation by modulating growth factor receptor signalling during the virus life cycle

    doi: 10.18632/oncotarget.21658

    Figure Lengend Snippet: AKT is essential for the modulation of keratinocyte differentiation by HPV18 (A) Schematic showing the KGFR signalling pathway and the effects of the dominant active (DA) and negative (DN) AKT. HPV18 genome containing keratinocytes were infected with empty retrovirus, or with retroviruses encoding DA AKT (B) or DN AKT (C) . Cells were differentiated in high calcium media for 48 hours prior to lysis and analysed for expression of cytokeratin 1 (K1) to assess differentiation. Expression of the exogenous AKT was confirmed using an antibody against the HA epitope and GAPDH served as a loading control.

    Article Snippet: Retrovirus transduction pLNCX AKT vectors (Addgene 9005, 9006 [ ]) were transfected into HEK 293T cells with murine retrovirus envelope and GAG/polymerase plasmids (kindly provided by Professor Greg Towers, University College London) using PEI transfection reagent.

    Techniques: Infection, Lysis, Expressing

    Cell spreading and directed migration is impaired in CD9/CD81-depleted MDA-MB-231. ( A ) MDA-MB-231 Wild type, CD9/CD81si, CD9RX, and CD81RX cells were plated on LM-332-coated glass coverslips, and cell spreading was imaged 30 min later by phase contrast microscopy. ( B ) The area of cell spreading for each cell type was calculated by subtracting the mean area of cells imaged immediately after plating from the mean area of cells after 30 min of spreading. Values are means ± s.e.m.; n = 3 trials with at least 25 cells of each cell type per trial; *P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: Cell spreading and directed migration is impaired in CD9/CD81-depleted MDA-MB-231. ( A ) MDA-MB-231 Wild type, CD9/CD81si, CD9RX, and CD81RX cells were plated on LM-332-coated glass coverslips, and cell spreading was imaged 30 min later by phase contrast microscopy. ( B ) The area of cell spreading for each cell type was calculated by subtracting the mean area of cells imaged immediately after plating from the mean area of cells after 30 min of spreading. Values are means ± s.e.m.; n = 3 trials with at least 25 cells of each cell type per trial; *P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Migration, Multiple Displacement Amplification, Microscopy

    CD151 promotes α3 integrin-CD9 association, and is required for normal adhesion and initial cell spreading on LM-332. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were lysed in 1% Brij 96V/Brij 99 (a 1∶1 mixture of both detergents), and CD9, CD81, CD151, or α3 integrin were immunoprecipitated (IPs), followed by blotting for the α3 integrin subunit. Note that α3 integrin-CD9 association is almost completely abolished in the CD151-silenced cells, but that α3-CD151 association is maintained in the CD9/CD81-silenced cells. ( B ) Wild type, CD9/CD81si, and CD151si MDA-MB-231 cells were plated in wells coated with LM-332, collagen I (COLI), or BSA for 30 min. Non-adherent cells were removed, and remaining cells were fixed and quantified by staining with crystal violet. Values are means ± s.e.m.; n = 4 wells/cell type. CD151si cells adhered less well than wild type or CD9/CD81si cells to LM-332 (*P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: CD151 promotes α3 integrin-CD9 association, and is required for normal adhesion and initial cell spreading on LM-332. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were lysed in 1% Brij 96V/Brij 99 (a 1∶1 mixture of both detergents), and CD9, CD81, CD151, or α3 integrin were immunoprecipitated (IPs), followed by blotting for the α3 integrin subunit. Note that α3 integrin-CD9 association is almost completely abolished in the CD151-silenced cells, but that α3-CD151 association is maintained in the CD9/CD81-silenced cells. ( B ) Wild type, CD9/CD81si, and CD151si MDA-MB-231 cells were plated in wells coated with LM-332, collagen I (COLI), or BSA for 30 min. Non-adherent cells were removed, and remaining cells were fixed and quantified by staining with crystal violet. Values are means ± s.e.m.; n = 4 wells/cell type. CD151si cells adhered less well than wild type or CD9/CD81si cells to LM-332 (*P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Multiple Displacement Amplification, Immunoprecipitation, Staining

    The CD9/CD81 complex promotes long-term growth of MDA-MB-231 cells in 3D Matrigel. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were suspended in growth factor reduced Matrigel and imaged at 7, 28, and 35 d time points, using a 20×, 10×, and 4× objective respectively. ( B ) Average colony growth was calculated by measuring the total area for 20–34 colonies per time point using ImageJ software. Bars indicate mean ± s.e.m. CD9/CD81si colonies, but not CD151si colonies, were significantly smaller than wild type colonies at the 28 and 35 d time points (*P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: The CD9/CD81 complex promotes long-term growth of MDA-MB-231 cells in 3D Matrigel. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were suspended in growth factor reduced Matrigel and imaged at 7, 28, and 35 d time points, using a 20×, 10×, and 4× objective respectively. ( B ) Average colony growth was calculated by measuring the total area for 20–34 colonies per time point using ImageJ software. Bars indicate mean ± s.e.m. CD9/CD81si colonies, but not CD151si colonies, were significantly smaller than wild type colonies at the 28 and 35 d time points (*P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Multiple Displacement Amplification, Software

    CD151 depletion does not impair MDA-MB-231 cell migration on LM-332. MDA-MB-231 wild type, CD9/CD81si, or CD151si cells were plated on LM-332 or collagen I-coated glass bottom dishes. After cells had attached and spread, motility was monitored by time-lapse microscopy, as in Fig. 1 . ( A ) CD9/CD81si cells, but not CD151si cells, displayed reduced migration velocity on LM-332 compared to wild type cells (*P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: CD151 depletion does not impair MDA-MB-231 cell migration on LM-332. MDA-MB-231 wild type, CD9/CD81si, or CD151si cells were plated on LM-332 or collagen I-coated glass bottom dishes. After cells had attached and spread, motility was monitored by time-lapse microscopy, as in Fig. 1 . ( A ) CD9/CD81si cells, but not CD151si cells, displayed reduced migration velocity on LM-332 compared to wild type cells (*P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Multiple Displacement Amplification, Migration, Time-lapse Microscopy

    CD9/CD81 promotes PKCα-α3β1 integrin association, and PKCα promotes α3β1-dependent MDA-MB-231 cell motility. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were treated with 100 nM PMA for 30 min, then lysed in 1% Brij 99 detergent followed by immunoprecipitation of CD9, CD151, α3 integrin, or CD55 and immunoblotting to detect PKCα. ( B,C ) Lysates of each cell type were immunoblotted for PKCα and β-actin. ( D ) The fraction of total cellular α3 integrin associated with PKCα in each cell type was estimated by LiCOR infrared fluorescence blot imaging, as described in Materials and Methods . Bars indicate mean ± S.E.M. from at least 3 different blots/cell type. CD9/CD81si cells showed a significant reduction in α3 integrin-associated PKCα compared to the other two cell types (*P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: CD9/CD81 promotes PKCα-α3β1 integrin association, and PKCα promotes α3β1-dependent MDA-MB-231 cell motility. ( A ) MDA-MB-231 wild type, CD9/CD81si, and CD151si cells were treated with 100 nM PMA for 30 min, then lysed in 1% Brij 99 detergent followed by immunoprecipitation of CD9, CD151, α3 integrin, or CD55 and immunoblotting to detect PKCα. ( B,C ) Lysates of each cell type were immunoblotted for PKCα and β-actin. ( D ) The fraction of total cellular α3 integrin associated with PKCα in each cell type was estimated by LiCOR infrared fluorescence blot imaging, as described in Materials and Methods . Bars indicate mean ± S.E.M. from at least 3 different blots/cell type. CD9/CD81si cells showed a significant reduction in α3 integrin-associated PKCα compared to the other two cell types (*P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Multiple Displacement Amplification, Immunoprecipitation, Fluorescence, Imaging

    Altered front-rear cell morphology in CD9/CD81-silenced cells. ( A ) Wild type MDA-MB-231 cells migrating on LM-332 frequently developed long protrusions at the rear of the cell (white arrows), which were less frequently observed in the CD9/CD81si cells. ( B ) The number of protrusions per cell lasting 4 min or longer during a 3 h video was quantified for wild type, CD9/CD81si, CD9RX, and CD81 RX cells. Values are means ± s.e.m.; n = 3 trials with 25 cells/trial. CD9/CD81si cells developed significantly fewer tails than wild type (*P

    Journal: PLoS ONE

    Article Title: The CD9/CD81 Tetraspanin Complex and Tetraspanin CD151 Regulate ?3?1 Integrin-Dependent Tumor Cell Behaviors by Overlapping but Distinct Mechanisms

    doi: 10.1371/journal.pone.0061834

    Figure Lengend Snippet: Altered front-rear cell morphology in CD9/CD81-silenced cells. ( A ) Wild type MDA-MB-231 cells migrating on LM-332 frequently developed long protrusions at the rear of the cell (white arrows), which were less frequently observed in the CD9/CD81si cells. ( B ) The number of protrusions per cell lasting 4 min or longer during a 3 h video was quantified for wild type, CD9/CD81si, CD9RX, and CD81 RX cells. Values are means ± s.e.m.; n = 3 trials with 25 cells/trial. CD9/CD81si cells developed significantly fewer tails than wild type (*P

    Article Snippet: Cell culture, RNAi and Retroviral Transduction MDA-MB-231 breast carcinoma (ATCC), A431 epithelial carcinoma (available from ATCC; obtained from the lab of Martin Hemler, Dana-Farber Cancer Institute) and GP2-293 retroviral packaging cells (Clontech) were cultured in high glucose DME medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/ml Penicillin, and 100 ug/ml Streptomycin (Invitrogen).

    Techniques: Multiple Displacement Amplification

    Autophagic flux in virus-specific CD8 T cells is inversely correlated with cell proliferation status. ( a ) MSCV retroviral constructs showing the transgenes (GFP-LC3b and GFP-LC3b-G120A) introduced in P14 cells to probe for autophagy activity. ( b ) Flow cytometry plots of adoptively transferred P14 cells in spleens. P14 cells transduced with retroviral plasmid of either MIT-GFP-LC3b or MIT-G120A are positive for the congenic marker Thy1.1. Day 5, 8 and 30 p.i. splenocytes were used for the analysis. The percentage of GFP-negative cells out of the transduced P14 (Thy1.1 + ) cells from each group is highlighted in blue on the lower right corner of each plot and is summarized in ( c ). ( d ) Representative ImageStream ® data showing images of P14 cells from either GFP-LC3b or G120A groups on day 8 p.i.. Images showing GFP signals of GFP + MIT vector-transduced P14 cells (Ly5.1 + Thy1.1 + ). Summary graph of ImageStream ® analysis is plotted in ( e ) showing percentage of GFP + or Thy1.1 + P14 cells that exhibited more than one GFP punctum. Errors bars represent SEM. ( b ) and ( d ) are representative of three independent experiments, n≥3 mice in each group.

    Journal: Nature immunology

    Article Title: Autophagy is essential for effector CD8 T cell survival and memory formation

    doi: 10.1038/ni.3025

    Figure Lengend Snippet: Autophagic flux in virus-specific CD8 T cells is inversely correlated with cell proliferation status. ( a ) MSCV retroviral constructs showing the transgenes (GFP-LC3b and GFP-LC3b-G120A) introduced in P14 cells to probe for autophagy activity. ( b ) Flow cytometry plots of adoptively transferred P14 cells in spleens. P14 cells transduced with retroviral plasmid of either MIT-GFP-LC3b or MIT-G120A are positive for the congenic marker Thy1.1. Day 5, 8 and 30 p.i. splenocytes were used for the analysis. The percentage of GFP-negative cells out of the transduced P14 (Thy1.1 + ) cells from each group is highlighted in blue on the lower right corner of each plot and is summarized in ( c ). ( d ) Representative ImageStream ® data showing images of P14 cells from either GFP-LC3b or G120A groups on day 8 p.i.. Images showing GFP signals of GFP + MIT vector-transduced P14 cells (Ly5.1 + Thy1.1 + ). Summary graph of ImageStream ® analysis is plotted in ( e ) showing percentage of GFP + or Thy1.1 + P14 cells that exhibited more than one GFP punctum. Errors bars represent SEM. ( b ) and ( d ) are representative of three independent experiments, n≥3 mice in each group.

    Article Snippet: Retroviral transduction Over-expression of transgenes in P14 was carried out using MSCV-IRES-Thy1.1 retrovirus vector (kindly provided by A. Rao, La Jolla Institute for Allergy and Immunology, La Jolla, CA; Addgene plasmid 17442).

    Techniques: Construct, Activity Assay, Flow Cytometry, Cytometry, Transduction, Plasmid Preparation, Marker, Mouse Assay