2x107 nuclei Search Results


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  • 99
    Covaris truchip chromatin shearing kit
    Truchip Chromatin Shearing Kit, supplied by Covaris, used in various techniques. Bioz Stars score: 99/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dtt
    Dtt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 31034 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad nitrocellulose membrane
    Nitrocellulose Membrane, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 46445 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher methanol free formaldehyde
    Methanol Free Formaldehyde, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 923 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega quantifluor dsdna system
    Quantifluor Dsdna System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 1997 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen ez1 platform
    Ez1 Platform, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Covaris protease inhibitors
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    Thermo Fisher superase in
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    Thermo Fisher rnase i peg10
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    Thermo Fisher lds lysis buffer
    Lds Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher paris kit
    Paris Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2862 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega m mlv reverse transcriptase
    M Mlv Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 33849 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rnase a
    Rnase A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher micrococcal nuclease
    Micrococcal Nuclease, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 999 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti mouse rae 1 antibody
    Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to <t>Rae-1</t> expressed in different tumor cells using flow cytometry.
    Anti Mouse Rae 1 Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche hiv 1 quantitative nucleic acid test
    Delayed viral rebound with 3BNC117 and 10-1074 combination therapy during ATI. a , Study design. Red and blue triangles represent 3BNC117 and 10-1074 infusions, respectively. b , Plasma <t>HIV-1</t> RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis) in the 9 bNAb-sensitive participants (left) and the 2 participants with pre-existing resistance against one of the antibodies (right). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. c , Kaplan-Meier plots summarizing time to viral rebound for the participants with HIV-1 RNA
    Hiv 1 Quantitative Nucleic Acid Test, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche protease inhibitors
    Delayed viral rebound with 3BNC117 and 10-1074 combination therapy during ATI. a , Study design. Red and blue triangles represent 3BNC117 and 10-1074 infusions, respectively. b , Plasma <t>HIV-1</t> RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis) in the 9 bNAb-sensitive participants (left) and the 2 participants with pre-existing resistance against one of the antibodies (right). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. c , Kaplan-Meier plots summarizing time to viral rebound for the participants with HIV-1 RNA
    Protease Inhibitors, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 81465 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore 8 bromo camp
    Intracellular <t>cAMP</t> controls nongenetic heterogeneity. (A) Growth-rate cumulative density curves of FY4 (black, 11900 microcolonies), FY4 cultivated with 15 mM <t>8-bromo-cAMP</t> (orange, 4510 microcolonies), ira2 (yellow, 6495 microcolonies) and pde2 (blue, 8666 microcolonies). Vertical axis is on a square-root scale for a better view of the slower-growing tail of each distribution. (B) Mean GFP fluorescence intensity—corrected by subtracting local background fluorescence then by subtracting the minimum value for the entire experiment, to avoid negative values (see Methods , vertical axis)—is plotted against microcolony growth rate (horizontal axis) for FY4 no-GFP control (black, 7340 microcolonies), TSL1-GFP (green, 6912 microcolonies), TSL1-GFP cultivated with 15 mM 8-bromo-cAMP (orange, 3730 microcolonies) and TSL1-GFP pde2 (blue, 1778 microcolonies). Each solid line is the fit to a generalized additive model with cubic spline smoother, with 95% confidence interval shown in yellow. Vertical axis is on a square-root scale for a better view at the low-intensity end.
    8 Bromo Camp, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 585 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher nuclear buffer
    Intracellular <t>cAMP</t> controls nongenetic heterogeneity. (A) Growth-rate cumulative density curves of FY4 (black, 11900 microcolonies), FY4 cultivated with 15 mM <t>8-bromo-cAMP</t> (orange, 4510 microcolonies), ira2 (yellow, 6495 microcolonies) and pde2 (blue, 8666 microcolonies). Vertical axis is on a square-root scale for a better view of the slower-growing tail of each distribution. (B) Mean GFP fluorescence intensity—corrected by subtracting local background fluorescence then by subtracting the minimum value for the entire experiment, to avoid negative values (see Methods , vertical axis)—is plotted against microcolony growth rate (horizontal axis) for FY4 no-GFP control (black, 7340 microcolonies), TSL1-GFP (green, 6912 microcolonies), TSL1-GFP cultivated with 15 mM 8-bromo-cAMP (orange, 3730 microcolonies) and TSL1-GFP pde2 (blue, 1778 microcolonies). Each solid line is the fit to a generalized additive model with cubic spline smoother, with 95% confidence interval shown in yellow. Vertical axis is on a square-root scale for a better view at the low-intensity end.
    Nuclear Buffer, supplied by Thermo Fisher, 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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    Roche lysis buffer
    Intracellular <t>cAMP</t> controls nongenetic heterogeneity. (A) Growth-rate cumulative density curves of FY4 (black, 11900 microcolonies), FY4 cultivated with 15 mM <t>8-bromo-cAMP</t> (orange, 4510 microcolonies), ira2 (yellow, 6495 microcolonies) and pde2 (blue, 8666 microcolonies). Vertical axis is on a square-root scale for a better view of the slower-growing tail of each distribution. (B) Mean GFP fluorescence intensity—corrected by subtracting local background fluorescence then by subtracting the minimum value for the entire experiment, to avoid negative values (see Methods , vertical axis)—is plotted against microcolony growth rate (horizontal axis) for FY4 no-GFP control (black, 7340 microcolonies), TSL1-GFP (green, 6912 microcolonies), TSL1-GFP cultivated with 15 mM 8-bromo-cAMP (orange, 3730 microcolonies) and TSL1-GFP pde2 (blue, 1778 microcolonies). Each solid line is the fit to a generalized additive model with cubic spline smoother, with 95% confidence interval shown in yellow. Vertical axis is on a square-root scale for a better view at the low-intensity end.
    Lysis Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 132608 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck KGaA benzonase nuclease
    KSHV LANA recruits Rad50 and Mre11 in the cytosol. ( A ) Co-immunoprecipitation of endogenous LANA, Rad50, Mre11 and Brd4 in BCBL-1 cells upon cytosolic-nuclear fractionation. Cells were lysed and cytoplasmic extracts (Cyto) and nuclear extracts (Nu) were prepared using the Thermo-Fischer Nu-Cyto fractionation kit following the manufacturer‘s instructions. Cytoplasmic and nuclear fractions were incubated overnight with sepharose beads coated with LANA-antibody or IgG-control. Left (INPUT, see Materials and methods ): Brd4, Lamin A/C and GAPDH immunoblots were analyzed to confirm the efficiency of the fractionation. Right (IP): immunoprecipitation with LANA-antibody or IgG-control coated-beads and immunoblot for endogenous Rad50, Mre11 and Brd4. ( B ) Co-immunoprecipitation of endogenous Rad50 and full-length LANA or ΔN mutants (Δ161 and Δ282) transfected into HEK293 cells. HEK293 cells were transfected with LANA constructs (or empty vector). 48 hours later cells were lysed and incubated with <t>benzonase.</t> After centrifugation, cells were incubated overnight with beads coated with LANA-antibody. Left (INPUT): immunoblot to check the expression of LANA constructs and the endogenous Rad50 in the cells. Right (IP from LANA-antibody-coated-beads): immunoblot for endogenous Rad50 co-immunoprecipitation. ( C ) Co-immunoprecipitation of endogenous LANA and Rad50, Mre11 and CARD9 in latently KSHV-infected THP-1 cells (TrK.219 cells, see Materials and methods ). Cells were lysed and incubated with benzonase. After centrifugation, whole cell lysates were incubated overnight with beads coated with anti-LANA or IgG-control antibody. Precipitated complexes were analysed by SDS-PAGE and immunoblotting with the indicated antibodies.
    Benzonase Nuclease, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 372 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche cobas ampliprep cobas taqman hiv 1 assay
    KSHV LANA recruits Rad50 and Mre11 in the cytosol. ( A ) Co-immunoprecipitation of endogenous LANA, Rad50, Mre11 and Brd4 in BCBL-1 cells upon cytosolic-nuclear fractionation. Cells were lysed and cytoplasmic extracts (Cyto) and nuclear extracts (Nu) were prepared using the Thermo-Fischer Nu-Cyto fractionation kit following the manufacturer‘s instructions. Cytoplasmic and nuclear fractions were incubated overnight with sepharose beads coated with LANA-antibody or IgG-control. Left (INPUT, see Materials and methods ): Brd4, Lamin A/C and GAPDH immunoblots were analyzed to confirm the efficiency of the fractionation. Right (IP): immunoprecipitation with LANA-antibody or IgG-control coated-beads and immunoblot for endogenous Rad50, Mre11 and Brd4. ( B ) Co-immunoprecipitation of endogenous Rad50 and full-length LANA or ΔN mutants (Δ161 and Δ282) transfected into HEK293 cells. HEK293 cells were transfected with LANA constructs (or empty vector). 48 hours later cells were lysed and incubated with <t>benzonase.</t> After centrifugation, cells were incubated overnight with beads coated with LANA-antibody. Left (INPUT): immunoblot to check the expression of LANA constructs and the endogenous Rad50 in the cells. Right (IP from LANA-antibody-coated-beads): immunoblot for endogenous Rad50 co-immunoprecipitation. ( C ) Co-immunoprecipitation of endogenous LANA and Rad50, Mre11 and CARD9 in latently KSHV-infected THP-1 cells (TrK.219 cells, see Materials and methods ). Cells were lysed and incubated with benzonase. After centrifugation, whole cell lysates were incubated overnight with beads coated with anti-LANA or IgG-control antibody. Precipitated complexes were analysed by SDS-PAGE and immunoblotting with the indicated antibodies.
    Cobas Ampliprep Cobas Taqman Hiv 1 Assay, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Becton Dickinson flow cytometry analysis
    Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow <t>cytometry.</t> Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).
    Flow Cytometry Analysis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 34287 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher propidium iodide
    Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow <t>cytometry.</t> Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).
    Propidium Iodide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 23878 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson in vitro flow cytometry analysis
    Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow <t>cytometry.</t> Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).
    In Vitro Flow Cytometry Analysis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Journal: PLoS ONE

    Article Title: Tumor-Targeted Delivery of IL-2 by NKG2D Leads to Accumulation of Antigen-Specific CD8+ T Cells in the Tumor Loci and Enhanced Anti-Tumor Effects

    doi: 10.1371/journal.pone.0035141

    Figure Lengend Snippet: Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Article Snippet: Flow Cytometry Analysis For in vitro flow cytometry analysis, samples of 2x105 tumor cells were incubated with 0.5μg purified recombinant protein or anti-mouse Rae-1 antibody (BD Bioscience).

    Techniques: Nucleic Acid Electrophoresis, Purification, Transfection, Construct, Isolation, SDS Page, Staining, Binding Assay, Flow Cytometry, Cytometry

    Delayed viral rebound with 3BNC117 and 10-1074 combination therapy during ATI. a , Study design. Red and blue triangles represent 3BNC117 and 10-1074 infusions, respectively. b , Plasma HIV-1 RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis) in the 9 bNAb-sensitive participants (left) and the 2 participants with pre-existing resistance against one of the antibodies (right). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. c , Kaplan-Meier plots summarizing time to viral rebound for the participants with HIV-1 RNA

    Journal: Nature

    Article Title: Combination therapy with anti-HIV-1 antibodies maintains viral suppression

    doi: 10.1038/s41586-018-0531-2

    Figure Lengend Snippet: Delayed viral rebound with 3BNC117 and 10-1074 combination therapy during ATI. a , Study design. Red and blue triangles represent 3BNC117 and 10-1074 infusions, respectively. b , Plasma HIV-1 RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis) in the 9 bNAb-sensitive participants (left) and the 2 participants with pre-existing resistance against one of the antibodies (right). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. c , Kaplan-Meier plots summarizing time to viral rebound for the participants with HIV-1 RNA

    Article Snippet: HIV-1 RNA levels were determined using the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Assay (version 2.0) or the Roche cobas HIV-1 quantitative nucleic acid test (cobas 6800), which quantitate HIV-1 RNA over a range of 2x101 to 1×107 copies/ml.

    Techniques:

    Viral rebound, amino acid variants at 10-1074 contact sites and sensitivities of latent and rebound viruses in the participants with detectable viremia > 20 copies/ml 2 weeks prior to or at the start of ATI. a , Plasma HIV-1 RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. b , Kaplan-Meier plots summarizing time to viral rebound. Y-axis indicates percentage of participants that maintain viral suppression. X-axis indicates weeks after start of ATI. Participants receiving the combination of 3BNC117 + 10-1074 are indicated by the blue line (n=4). Dotted red line indicates a cohort of individuals receiving 3BNC117 alone during ATI 9 (n=13) and dotted black line indicates a cohort of participants who underwent ATI without any intervention 10 (n=52). c , Color charts show Env contact sites of 10-1074 at the G(D/N)IR motif (positions 324-327, according to HXB2 numbering) and the glycan at the potential N -linked glycosylation site at position 332 (NxS/T motif at positions 332-334). LR indicates latent reservoir viruses isolated by Q 2 VOA (week -2) and RB indicates rebound viruses isolated by SGA (plasma) or viral outgrowth (PBMCs). Each amino acid is represented by a color and the frequency of each amino acid is indicated by the height of the rectangle. Shaded rectangles indicate the lack of variation between latent reservoir virus and rebound virus at the indicated position. Full-color rectangles represent amino acid residues with changes in distribution between reservoir and rebound viruses. d , Dot plots indicating IC 80 (µg/ml) of 3BNC117 (left) and 10-1074 (right) against latent and rebound viruses determined by TZM-bl neutralization assay. Q 2 VOA-derived latent viruses from week -2 are shown as black circles. For outgrowth culture-derived rebound viruses, the highest IC 80 determined is shown as red circle. For 9250 and 9253, no viruses could be obtained from rebound outgrowth cultures and pseudoviruses were made from env sequences of the latent reservoir (Q 2 VOA) and rebound viruses (plasma SGA). Note that 9249 and 9253 had pre-existing resistant viruses in the reservoir (IC 50 > 2 μg/ml). 9248 and 9250 had pre-existing viruses that failed to reach an IC 100 when tested up to 50 µg/ml for 3BNC117 ( Extended Data Fig. 5 ). Rebound viruses of all 4 participants showed IC 80 or IC 100 of > 50 μg/ml fo r both 3BN117 and 10-1074.

    Journal: Nature

    Article Title: Combination therapy with anti-HIV-1 antibodies maintains viral suppression

    doi: 10.1038/s41586-018-0531-2

    Figure Lengend Snippet: Viral rebound, amino acid variants at 10-1074 contact sites and sensitivities of latent and rebound viruses in the participants with detectable viremia > 20 copies/ml 2 weeks prior to or at the start of ATI. a , Plasma HIV-1 RNA levels (black; left y-axis) and bNAb serum concentrations (3BNC117, red; 10-1074, blue; right y-axis). Red and blue triangles indicate 3BNC117 and 10-1074 infusions, respectively. Serum antibody concentrations were determined by TZM-bl assay. Grey shaded areas indicate time on ART. Lower limit of detection of HIV-1 RNA was 20 copies/ml. b , Kaplan-Meier plots summarizing time to viral rebound. Y-axis indicates percentage of participants that maintain viral suppression. X-axis indicates weeks after start of ATI. Participants receiving the combination of 3BNC117 + 10-1074 are indicated by the blue line (n=4). Dotted red line indicates a cohort of individuals receiving 3BNC117 alone during ATI 9 (n=13) and dotted black line indicates a cohort of participants who underwent ATI without any intervention 10 (n=52). c , Color charts show Env contact sites of 10-1074 at the G(D/N)IR motif (positions 324-327, according to HXB2 numbering) and the glycan at the potential N -linked glycosylation site at position 332 (NxS/T motif at positions 332-334). LR indicates latent reservoir viruses isolated by Q 2 VOA (week -2) and RB indicates rebound viruses isolated by SGA (plasma) or viral outgrowth (PBMCs). Each amino acid is represented by a color and the frequency of each amino acid is indicated by the height of the rectangle. Shaded rectangles indicate the lack of variation between latent reservoir virus and rebound virus at the indicated position. Full-color rectangles represent amino acid residues with changes in distribution between reservoir and rebound viruses. d , Dot plots indicating IC 80 (µg/ml) of 3BNC117 (left) and 10-1074 (right) against latent and rebound viruses determined by TZM-bl neutralization assay. Q 2 VOA-derived latent viruses from week -2 are shown as black circles. For outgrowth culture-derived rebound viruses, the highest IC 80 determined is shown as red circle. For 9250 and 9253, no viruses could be obtained from rebound outgrowth cultures and pseudoviruses were made from env sequences of the latent reservoir (Q 2 VOA) and rebound viruses (plasma SGA). Note that 9249 and 9253 had pre-existing resistant viruses in the reservoir (IC 50 > 2 μg/ml). 9248 and 9250 had pre-existing viruses that failed to reach an IC 100 when tested up to 50 µg/ml for 3BNC117 ( Extended Data Fig. 5 ). Rebound viruses of all 4 participants showed IC 80 or IC 100 of > 50 μg/ml fo r both 3BN117 and 10-1074.

    Article Snippet: HIV-1 RNA levels were determined using the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Assay (version 2.0) or the Roche cobas HIV-1 quantitative nucleic acid test (cobas 6800), which quantitate HIV-1 RNA over a range of 2x101 to 1×107 copies/ml.

    Techniques: Isolation, Neutralization, Derivative Assay

    Intracellular cAMP controls nongenetic heterogeneity. (A) Growth-rate cumulative density curves of FY4 (black, 11900 microcolonies), FY4 cultivated with 15 mM 8-bromo-cAMP (orange, 4510 microcolonies), ira2 (yellow, 6495 microcolonies) and pde2 (blue, 8666 microcolonies). Vertical axis is on a square-root scale for a better view of the slower-growing tail of each distribution. (B) Mean GFP fluorescence intensity—corrected by subtracting local background fluorescence then by subtracting the minimum value for the entire experiment, to avoid negative values (see Methods , vertical axis)—is plotted against microcolony growth rate (horizontal axis) for FY4 no-GFP control (black, 7340 microcolonies), TSL1-GFP (green, 6912 microcolonies), TSL1-GFP cultivated with 15 mM 8-bromo-cAMP (orange, 3730 microcolonies) and TSL1-GFP pde2 (blue, 1778 microcolonies). Each solid line is the fit to a generalized additive model with cubic spline smoother, with 95% confidence interval shown in yellow. Vertical axis is on a square-root scale for a better view at the low-intensity end.

    Journal: PLoS Genetics

    Article Title: Control of nongenetic heterogeneity in growth rate and stress tolerance of Saccharomyces cerevisiae by cyclic AMP-regulated transcription factors

    doi: 10.1371/journal.pgen.1007744

    Figure Lengend Snippet: Intracellular cAMP controls nongenetic heterogeneity. (A) Growth-rate cumulative density curves of FY4 (black, 11900 microcolonies), FY4 cultivated with 15 mM 8-bromo-cAMP (orange, 4510 microcolonies), ira2 (yellow, 6495 microcolonies) and pde2 (blue, 8666 microcolonies). Vertical axis is on a square-root scale for a better view of the slower-growing tail of each distribution. (B) Mean GFP fluorescence intensity—corrected by subtracting local background fluorescence then by subtracting the minimum value for the entire experiment, to avoid negative values (see Methods , vertical axis)—is plotted against microcolony growth rate (horizontal axis) for FY4 no-GFP control (black, 7340 microcolonies), TSL1-GFP (green, 6912 microcolonies), TSL1-GFP cultivated with 15 mM 8-bromo-cAMP (orange, 3730 microcolonies) and TSL1-GFP pde2 (blue, 1778 microcolonies). Each solid line is the fit to a generalized additive model with cubic spline smoother, with 95% confidence interval shown in yellow. Vertical axis is on a square-root scale for a better view at the low-intensity end.

    Article Snippet: For addition of the cAMP analog, fresh SC liquid medium was supplemented with 15 mM 8-bromo-cAMP (Sigma, Catalog Number: B7880, dissolved in water) and culture tubes were wrapped with foil to protect the light-sensitive compound. ira2 and bcy1 mutants were diluted to about 2X104 cells/ml into fresh SC liquid media.

    Techniques: Fluorescence

    Perturbations of the Ras/cAMP/PKA/Msn2/4 pathway affect acute heat-stress tolerance. (A) Rate of survival of acute heat shock (51° C for 2 min) for FY4, msn2 , msn4 , msn2 msn4 , pde2 , ira2 and FY4 treated with 15 mM 8-bromo cAMP. Star symbols indicate the mean. Error bars indicate standard error of the mean. Individual data points (three replicates per strain) are shown to the right of each mean-and-error bar. Statistical significance is indicated for pairwise comparisons (*** = adjusted P

    Journal: PLoS Genetics

    Article Title: Control of nongenetic heterogeneity in growth rate and stress tolerance of Saccharomyces cerevisiae by cyclic AMP-regulated transcription factors

    doi: 10.1371/journal.pgen.1007744

    Figure Lengend Snippet: Perturbations of the Ras/cAMP/PKA/Msn2/4 pathway affect acute heat-stress tolerance. (A) Rate of survival of acute heat shock (51° C for 2 min) for FY4, msn2 , msn4 , msn2 msn4 , pde2 , ira2 and FY4 treated with 15 mM 8-bromo cAMP. Star symbols indicate the mean. Error bars indicate standard error of the mean. Individual data points (three replicates per strain) are shown to the right of each mean-and-error bar. Statistical significance is indicated for pairwise comparisons (*** = adjusted P

    Article Snippet: For addition of the cAMP analog, fresh SC liquid medium was supplemented with 15 mM 8-bromo-cAMP (Sigma, Catalog Number: B7880, dissolved in water) and culture tubes were wrapped with foil to protect the light-sensitive compound. ira2 and bcy1 mutants were diluted to about 2X104 cells/ml into fresh SC liquid media.

    Techniques:

    KSHV LANA recruits Rad50 and Mre11 in the cytosol. ( A ) Co-immunoprecipitation of endogenous LANA, Rad50, Mre11 and Brd4 in BCBL-1 cells upon cytosolic-nuclear fractionation. Cells were lysed and cytoplasmic extracts (Cyto) and nuclear extracts (Nu) were prepared using the Thermo-Fischer Nu-Cyto fractionation kit following the manufacturer‘s instructions. Cytoplasmic and nuclear fractions were incubated overnight with sepharose beads coated with LANA-antibody or IgG-control. Left (INPUT, see Materials and methods ): Brd4, Lamin A/C and GAPDH immunoblots were analyzed to confirm the efficiency of the fractionation. Right (IP): immunoprecipitation with LANA-antibody or IgG-control coated-beads and immunoblot for endogenous Rad50, Mre11 and Brd4. ( B ) Co-immunoprecipitation of endogenous Rad50 and full-length LANA or ΔN mutants (Δ161 and Δ282) transfected into HEK293 cells. HEK293 cells were transfected with LANA constructs (or empty vector). 48 hours later cells were lysed and incubated with benzonase. After centrifugation, cells were incubated overnight with beads coated with LANA-antibody. Left (INPUT): immunoblot to check the expression of LANA constructs and the endogenous Rad50 in the cells. Right (IP from LANA-antibody-coated-beads): immunoblot for endogenous Rad50 co-immunoprecipitation. ( C ) Co-immunoprecipitation of endogenous LANA and Rad50, Mre11 and CARD9 in latently KSHV-infected THP-1 cells (TrK.219 cells, see Materials and methods ). Cells were lysed and incubated with benzonase. After centrifugation, whole cell lysates were incubated overnight with beads coated with anti-LANA or IgG-control antibody. Precipitated complexes were analysed by SDS-PAGE and immunoblotting with the indicated antibodies.

    Journal: PLoS Pathogens

    Article Title: Kaposi Sarcoma Herpesvirus (KSHV) Latency-Associated Nuclear Antigen (LANA) recruits components of the MRN (Mre11-Rad50-NBS1) repair complex to modulate an innate immune signaling pathway and viral latency

    doi: 10.1371/journal.ppat.1006335

    Figure Lengend Snippet: KSHV LANA recruits Rad50 and Mre11 in the cytosol. ( A ) Co-immunoprecipitation of endogenous LANA, Rad50, Mre11 and Brd4 in BCBL-1 cells upon cytosolic-nuclear fractionation. Cells were lysed and cytoplasmic extracts (Cyto) and nuclear extracts (Nu) were prepared using the Thermo-Fischer Nu-Cyto fractionation kit following the manufacturer‘s instructions. Cytoplasmic and nuclear fractions were incubated overnight with sepharose beads coated with LANA-antibody or IgG-control. Left (INPUT, see Materials and methods ): Brd4, Lamin A/C and GAPDH immunoblots were analyzed to confirm the efficiency of the fractionation. Right (IP): immunoprecipitation with LANA-antibody or IgG-control coated-beads and immunoblot for endogenous Rad50, Mre11 and Brd4. ( B ) Co-immunoprecipitation of endogenous Rad50 and full-length LANA or ΔN mutants (Δ161 and Δ282) transfected into HEK293 cells. HEK293 cells were transfected with LANA constructs (or empty vector). 48 hours later cells were lysed and incubated with benzonase. After centrifugation, cells were incubated overnight with beads coated with LANA-antibody. Left (INPUT): immunoblot to check the expression of LANA constructs and the endogenous Rad50 in the cells. Right (IP from LANA-antibody-coated-beads): immunoblot for endogenous Rad50 co-immunoprecipitation. ( C ) Co-immunoprecipitation of endogenous LANA and Rad50, Mre11 and CARD9 in latently KSHV-infected THP-1 cells (TrK.219 cells, see Materials and methods ). Cells were lysed and incubated with benzonase. After centrifugation, whole cell lysates were incubated overnight with beads coated with anti-LANA or IgG-control antibody. Precipitated complexes were analysed by SDS-PAGE and immunoblotting with the indicated antibodies.

    Article Snippet: Benzonase nuclease (Merck Millipore, 71205–3) was added to whole cell lysates (50U each 2x106 cells) for 30 minutes at RT to digest nucleic acids.

    Techniques: Immunoprecipitation, Fractionation, Incubation, Western Blot, Transfection, Construct, Plasmid Preparation, Centrifugation, Expressing, Infection, SDS Page

    KSHV LANA recruits MRN (Mre11-Rad50-NBS1) complex. ( A ) Co-immunoprecipitation of endogenous LANA and MRN proteins in BC3 cells. Cells were lysed using TBS-T buffer and the cell lysate was incubated with benzonase. After centrifugation, supernatant was incubated overnight with anti-LANA or IgG-control beads. The precipitated complexes were analyzed for the presence of endogenous Rad50, Mre11 and NBS1 by SDS-PAGE and immunoblotting. For the input, see Materials and methods . ( B ) Co-immunoprecipitation of endogenous LANA and Rad50 in BC3 cells. Co-immunoprecipitation of endogenous Rad50 was performed and analysed as in (A), but with anti-Rad50-antibody-coated-beads (left) or anti-LANA coated beads (right). The arrowhead indicates the smaller LANA forms co-immunoprecipitating with Rad50 (see text). ( C ) Schematic representation of LANA domain structure. NLS: Nuclear Localization Signal; TR: KSHV Terminal Repeats. ( D ) Pull-down assay with GST-fused LANA-C (aa 931–1162) and LANA-N (aa 1–312) proteins and HEK293T cell lysates. HEK293T were lysed with TBS-T buffer and incubated 4 hours with GST-fused proteins or GST alone, as negative control. Top : immunoblot for endogenous Rad50, Mre11 and NBS1 bound to GST-fused LANA fragments. Bottom : Ponceau staining to detect GST-fused proteins. (M) for marker.

    Journal: PLoS Pathogens

    Article Title: Kaposi Sarcoma Herpesvirus (KSHV) Latency-Associated Nuclear Antigen (LANA) recruits components of the MRN (Mre11-Rad50-NBS1) repair complex to modulate an innate immune signaling pathway and viral latency

    doi: 10.1371/journal.ppat.1006335

    Figure Lengend Snippet: KSHV LANA recruits MRN (Mre11-Rad50-NBS1) complex. ( A ) Co-immunoprecipitation of endogenous LANA and MRN proteins in BC3 cells. Cells were lysed using TBS-T buffer and the cell lysate was incubated with benzonase. After centrifugation, supernatant was incubated overnight with anti-LANA or IgG-control beads. The precipitated complexes were analyzed for the presence of endogenous Rad50, Mre11 and NBS1 by SDS-PAGE and immunoblotting. For the input, see Materials and methods . ( B ) Co-immunoprecipitation of endogenous LANA and Rad50 in BC3 cells. Co-immunoprecipitation of endogenous Rad50 was performed and analysed as in (A), but with anti-Rad50-antibody-coated-beads (left) or anti-LANA coated beads (right). The arrowhead indicates the smaller LANA forms co-immunoprecipitating with Rad50 (see text). ( C ) Schematic representation of LANA domain structure. NLS: Nuclear Localization Signal; TR: KSHV Terminal Repeats. ( D ) Pull-down assay with GST-fused LANA-C (aa 931–1162) and LANA-N (aa 1–312) proteins and HEK293T cell lysates. HEK293T were lysed with TBS-T buffer and incubated 4 hours with GST-fused proteins or GST alone, as negative control. Top : immunoblot for endogenous Rad50, Mre11 and NBS1 bound to GST-fused LANA fragments. Bottom : Ponceau staining to detect GST-fused proteins. (M) for marker.

    Article Snippet: Benzonase nuclease (Merck Millipore, 71205–3) was added to whole cell lysates (50U each 2x106 cells) for 30 minutes at RT to digest nucleic acids.

    Techniques: Immunoprecipitation, Incubation, Centrifugation, SDS Page, Pull Down Assay, Negative Control, Staining, Marker

    Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow cytometry. Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).

    Journal: PLoS ONE

    Article Title: Tumor-Targeted Delivery of IL-2 by NKG2D Leads to Accumulation of Antigen-Specific CD8+ T Cells in the Tumor Loci and Enhanced Anti-Tumor Effects

    doi: 10.1371/journal.pone.0035141

    Figure Lengend Snippet: Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow cytometry. Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).

    Article Snippet: Flow Cytometry Analysis For in vitro flow cytometry analysis, samples of 2x105 tumor cells were incubated with 0.5μg purified recombinant protein or anti-mouse Rae-1 antibody (BD Bioscience).

    Techniques: Mouse Assay, Injection, Construct, Electroporation, Binding Assay, Incubation, Flow Cytometry, Cytometry, Imaging, Luciferase, Expressing, Significance Assay

    Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Journal: PLoS ONE

    Article Title: Tumor-Targeted Delivery of IL-2 by NKG2D Leads to Accumulation of Antigen-Specific CD8+ T Cells in the Tumor Loci and Enhanced Anti-Tumor Effects

    doi: 10.1371/journal.pone.0035141

    Figure Lengend Snippet: Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Article Snippet: Flow Cytometry Analysis For in vitro flow cytometry analysis, samples of 2x105 tumor cells were incubated with 0.5μg purified recombinant protein or anti-mouse Rae-1 antibody (BD Bioscience).

    Techniques: Nucleic Acid Electrophoresis, Purification, Transfection, Construct, Isolation, SDS Page, Staining, Binding Assay, Flow Cytometry, Cytometry

    Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow cytometry. Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).

    Journal: PLoS ONE

    Article Title: Tumor-Targeted Delivery of IL-2 by NKG2D Leads to Accumulation of Antigen-Specific CD8+ T Cells in the Tumor Loci and Enhanced Anti-Tumor Effects

    doi: 10.1371/journal.pone.0035141

    Figure Lengend Snippet: Characterization of T cell accumulation in tumor-bearing mice intramuscularly injected with DNA constructs with electroporation. (A) Characterization of the binding of NKG2D-Fc-containing protein to TC-1 tumor cell using serum from mice injected intramuscularly with the various DNA constructs in conjunction with electroporation. Mice were intramuscularly injected with one of four DNA constructs. Serum was collected from the mice and incubated with TC-1 tumor cells. The cells were incubated with anti-Fc-PE and characterized by flow cytometry. Note: the tumor cells incubated with the serum from mice injected with the DNA constructs encoding NKG2D-Fc or NKG2D-Fc-IL2 led to a significant shift compared to other constructs. (B) Representative bioluminescence imaging used to characterize the presence of luciferase-expressing E7-specific CD8+ T cells in tumor-bearing mice injected intramuscularly with different DNA constructs followed by electroporation. TC-1 tumor bearing mice were injected intramuscularly with one of the four DNA constructs, followed by electroporation. Luciferase-expressing T cells were injected intravenously into the lateral tail vein and the bioluminescence was imaged five days later. Note: the luciferase-expressing T cells proliferated locally to the tumor site in the mice injected with DNA encoding NKG2D-Fc-IL2 but not in others. (C) Histogram of the bioluminescence imaging depicted in Figure 4B . Note: the luminescence of the tumors between the mice injected with NKG2D-Fc-IL2 and those not (P-value less than 0.001).

    Article Snippet: Flow Cytometry Analysis For in vitro flow cytometry analysis, samples of 2x105 tumor cells were incubated with 0.5μg purified recombinant protein or anti-mouse Rae-1 antibody (BD Bioscience).

    Techniques: Mouse Assay, Injection, Construct, Electroporation, Binding Assay, Incubation, Flow Cytometry, Cytometry, Imaging, Luciferase, Expressing, Significance Assay

    Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Journal: PLoS ONE

    Article Title: Tumor-Targeted Delivery of IL-2 by NKG2D Leads to Accumulation of Antigen-Specific CD8+ T Cells in the Tumor Loci and Enhanced Anti-Tumor Effects

    doi: 10.1371/journal.pone.0035141

    Figure Lengend Snippet: Characterization of the murine NKG2D-Fc protein. (A) Gel electrophoresis was used to characterize the size of the purified NKG2D-Fc and Con-Fc proteins. The NKG2D-Fc and Con-Fc proteins were purified from BHK-21 cells transfected with the pFuse-Fc and pFuse-NKG2D-Fc DNA constructs. The purity and size of isolated protein was characterized by SDS-PAGE, followed by staining with Coomassie brilliant blue. The arrows indicate the proteins of interest. (B) Characterization of the binding of NKG2D-Fc to Rae-1 expressed in different tumor cells using flow cytometry.

    Article Snippet: Flow Cytometry Analysis For in vitro flow cytometry analysis, samples of 2x105 tumor cells were incubated with 0.5μg purified recombinant protein or anti-mouse Rae-1 antibody (BD Bioscience).

    Techniques: Nucleic Acid Electrophoresis, Purification, Transfection, Construct, Isolation, SDS Page, Staining, Binding Assay, Flow Cytometry, Cytometry