trypsin-edta solution Search Results


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  • 99
    Lonza trypsin edta solution
    Trypsin Edta Solution, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 399 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC trypsin edta solution
    Trypsin Edta Solution, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher trypsin edta
    The suppression of OCs on T-cell proliferation is IFN-γ and CD40L dependent Proliferation of CD4 + T cells stimulated by (A) α-CD3/CD28 Dynabeads at a T/Bead ratio of 2:1 or (B) allogeneic DCs at a T/DC ratio of 10:1 in <t>Transwells</t> in the absence or presence of autologous OCs with IFN-γ neutralizing or IFNGR1 blocking antibodies (20 μg/mL) for 4–6 d. (C) Proliferation of CD4 + T cells stimulated by allogeneic DCs under the contact coculture of autologous OCs with IFN-γ neutralizing, CD40 or CD40L blocking antibodies (20 μg/mL). (D) Expression of IFNGR1 and CD40 on OCs, as measured by flow cytometry. OCs coculutred with α-CD3/CD28 Dynabeads-activated CD4 + T cells in Transwell system were harvested by <t>Trypsin/EDTA</t> detachment, stained by antibodies and analyzed by flow cytometry. Representative results from 3 independent experiments are shown.
    Trypsin Edta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 52239 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 3526 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore trypsin edta
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Biochrom trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Biochrom, used in various techniques. Bioz Stars score: 92/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Beyotime trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore trypsin ethylenediaminetetraacetic acid edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Ethylenediaminetetraacetic Acid Edta Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    GE Healthcare trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 98/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Mediatech trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Mediatech, used in various techniques. Bioz Stars score: 92/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nacalai trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Nacalai, used in various techniques. Bioz Stars score: 93/100, based on 144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PAA Laboratories trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biological Industries Inc trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Biological Industries Inc, used in various techniques. Bioz Stars score: 93/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Welgene inc trypsin edta solution
    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in <t>PBS</t> (columns 1, 3, and 5) or treated with a <t>trypsin-EDTA</t> solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.
    Trypsin Edta Solution, supplied by Welgene inc, used in various techniques. Bioz Stars score: 90/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The suppression of OCs on T-cell proliferation is IFN-γ and CD40L dependent Proliferation of CD4 + T cells stimulated by (A) α-CD3/CD28 Dynabeads at a T/Bead ratio of 2:1 or (B) allogeneic DCs at a T/DC ratio of 10:1 in Transwells in the absence or presence of autologous OCs with IFN-γ neutralizing or IFNGR1 blocking antibodies (20 μg/mL) for 4–6 d. (C) Proliferation of CD4 + T cells stimulated by allogeneic DCs under the contact coculture of autologous OCs with IFN-γ neutralizing, CD40 or CD40L blocking antibodies (20 μg/mL). (D) Expression of IFNGR1 and CD40 on OCs, as measured by flow cytometry. OCs coculutred with α-CD3/CD28 Dynabeads-activated CD4 + T cells in Transwell system were harvested by Trypsin/EDTA detachment, stained by antibodies and analyzed by flow cytometry. Representative results from 3 independent experiments are shown.

    Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

    Article Title: Human Osteoclasts are Inducible Immunosuppressive Cells in Response to T cell-Derived IFN-γ and CD40 Ligand in vitro

    doi: 10.1002/jbmr.2294

    Figure Lengend Snippet: The suppression of OCs on T-cell proliferation is IFN-γ and CD40L dependent Proliferation of CD4 + T cells stimulated by (A) α-CD3/CD28 Dynabeads at a T/Bead ratio of 2:1 or (B) allogeneic DCs at a T/DC ratio of 10:1 in Transwells in the absence or presence of autologous OCs with IFN-γ neutralizing or IFNGR1 blocking antibodies (20 μg/mL) for 4–6 d. (C) Proliferation of CD4 + T cells stimulated by allogeneic DCs under the contact coculture of autologous OCs with IFN-γ neutralizing, CD40 or CD40L blocking antibodies (20 μg/mL). (D) Expression of IFNGR1 and CD40 on OCs, as measured by flow cytometry. OCs coculutred with α-CD3/CD28 Dynabeads-activated CD4 + T cells in Transwell system were harvested by Trypsin/EDTA detachment, stained by antibodies and analyzed by flow cytometry. Representative results from 3 independent experiments are shown.

    Article Snippet: To detect the viability of OCs during the coculture with α-CD3/CD28 DynaBeads-stimulated T cells in Transwells, coculutred OCs were digested from the culture plate with Trypsin/EDTA, and stained by using LIVE/DEAD® Fixable Dead Cell Stain Kit (Molecular Probes, Life Technologies).

    Techniques: Blocking Assay, Expressing, Flow Cytometry, Cytometry, Staining

    Sevoflurane and rat brain endothelial cell size in H/R injury. RBE4 cells were exposed to severe hypoxia (0.2% oxygen) for 24 hours, followed by a 4-hour period of reoxygenation in air (H/R+air) or air enriched with sevoflurane (H/R+sevo). Cells in the normoxia group remained in a normal cell culture environment with 21% oxygen for the full 28 hours. After detachment with trypsin-EDTA and viability staining, cells were analyzed with the flow cytometer or image stream, respectively. FSC-A as an indirect measure of cell size was assessed in conventional flow cytometry ( A ), while the cell size of each cell was assessed with the image stream method ( B ). The histogram shows the distribution pattern of FSC-A seen in the flow cytometry. n = 5 independent experiments, with 500 cells analyzed in each condition. The bar graph shows mean values and standard deviations. n = 4 Image stream X experiments, at least 20,000 cells analyzed in each condition. Analysis with one way ANOVA and Bonferroni correction. *** p

    Journal: PLoS ONE

    Article Title: Sevoflurane protects rat brain endothelial barrier structure and function after hypoxia-reoxygenation injury

    doi: 10.1371/journal.pone.0184973

    Figure Lengend Snippet: Sevoflurane and rat brain endothelial cell size in H/R injury. RBE4 cells were exposed to severe hypoxia (0.2% oxygen) for 24 hours, followed by a 4-hour period of reoxygenation in air (H/R+air) or air enriched with sevoflurane (H/R+sevo). Cells in the normoxia group remained in a normal cell culture environment with 21% oxygen for the full 28 hours. After detachment with trypsin-EDTA and viability staining, cells were analyzed with the flow cytometer or image stream, respectively. FSC-A as an indirect measure of cell size was assessed in conventional flow cytometry ( A ), while the cell size of each cell was assessed with the image stream method ( B ). The histogram shows the distribution pattern of FSC-A seen in the flow cytometry. n = 5 independent experiments, with 500 cells analyzed in each condition. The bar graph shows mean values and standard deviations. n = 4 Image stream X experiments, at least 20,000 cells analyzed in each condition. Analysis with one way ANOVA and Bonferroni correction. *** p

    Article Snippet: Flow cytometry and cell size measurement For flow and imaging cytometry cells were washed twice with 2.5mmol EDTA in PBS, detached with trypsin-EDTA solution (0.05%) (Thermo, Reinach, Switzerland) and put through a 35μm cell strainer (Falcon, Corning, USA) after resuspension.

    Techniques: Cell Culture, Staining, Flow Cytometry, Cytometry

    Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in PBS (columns 1, 3, and 5) or treated with a trypsin-EDTA solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.

    Journal: Journal of Virology

    Article Title: Compensatory Link between Fusion and Endocytosis of Human Immunodeficiency Virus Type 1 in Human CD4 T Lymphocytes

    doi: 10.1128/JVI.78.3.1375-1383.2004

    Figure Lengend Snippet: Both surface binding and intracellular entry by fusion of HIV-1 virions are enhanced by endosome inhibitors. SupT1 cells were pretreated for 30 min at 37°C with medium (columns 1 and 2), bafilomycin A1 (100 nM; columns 3 and 4), or NH 4 Cl (10 mM; columns 5 and 6). Cells were then incubated with GFP-Vpr-labeled HIV-1 virions (200 ng of p24 Gag) for 3 h at 37°C. Cells were either washed in PBS (columns 1, 3, and 5) or treated with a trypsin-EDTA solution to remove surface-bound virions (columns 2, 4, and 6) followed by flow cytometric analysis of living cells. The percentage of SupT1 cells displaying GFP epifluorescence is indicated on the ordinate. The data presented are from a representative experiment repeated three times with comparable results.

    Article Snippet: Cells were washed with phosphate-buffered saline (PBS) or treated with a trypsin-EDTA solution (1×; Sigma) for 2 min at room temperature to remove surface-bound virions, followed by additional washing and fixation in 1% paraformaldehyde-PBS solution.

    Techniques: Binding Assay, Incubation, Labeling, Flow Cytometry

    Flow cytometric analysis of PHA-activated human lymphoblasts infected with GFP-Vpr-labeled HIV-1 virions. (A) PBMCs activated with PHA and cultured in interleukin-2 for 4 days were inoculated with GFP-Vpr-labeled X4-tropic HIV-1 (300 ng of p24 Gag) at 37 or 4°C as indicated. After 3 h, the cells were washed at room temperature with PBS (−T) or a trypsin-EDTA solution (+T) to remove surface-bound virions. Live cells were then analyzed by flow cytometry to detect GFP epifluorescence. Error bars indicate standard deviations of the mean derived from two independent experiments. (B and C) PHA-activated human lymphoblasts were pretreated for 30 min at 37°C with either medium (panels 2 and 4) or AMD3100 (panels 3 and 5). Cells were then incubated at 37°C for 3 or 24 h with GFP-Vpr-labeled X4-tropic HIV-1 or HIV virions pseudotyped with the VSV-G envelope (200 ng of p24 Gag) at 37°C. Cells were subsequently stained with APC-conjugated CD4 antibodies (B) or PE-conjugated CD4 antibodies (C). Cells were analyzed for GFP epifluorescence and APC or PE immunofluorescence. The percentage of cells in each quadrant is indicated. Data shown are from a representative experiment performed three times with comparable results. Note preferential entry of HIV into CD4 cells and the absence of inhibitory effects of AMD3100 measured either at 3 or 24 h.

    Journal: Journal of Virology

    Article Title: Compensatory Link between Fusion and Endocytosis of Human Immunodeficiency Virus Type 1 in Human CD4 T Lymphocytes

    doi: 10.1128/JVI.78.3.1375-1383.2004

    Figure Lengend Snippet: Flow cytometric analysis of PHA-activated human lymphoblasts infected with GFP-Vpr-labeled HIV-1 virions. (A) PBMCs activated with PHA and cultured in interleukin-2 for 4 days were inoculated with GFP-Vpr-labeled X4-tropic HIV-1 (300 ng of p24 Gag) at 37 or 4°C as indicated. After 3 h, the cells were washed at room temperature with PBS (−T) or a trypsin-EDTA solution (+T) to remove surface-bound virions. Live cells were then analyzed by flow cytometry to detect GFP epifluorescence. Error bars indicate standard deviations of the mean derived from two independent experiments. (B and C) PHA-activated human lymphoblasts were pretreated for 30 min at 37°C with either medium (panels 2 and 4) or AMD3100 (panels 3 and 5). Cells were then incubated at 37°C for 3 or 24 h with GFP-Vpr-labeled X4-tropic HIV-1 or HIV virions pseudotyped with the VSV-G envelope (200 ng of p24 Gag) at 37°C. Cells were subsequently stained with APC-conjugated CD4 antibodies (B) or PE-conjugated CD4 antibodies (C). Cells were analyzed for GFP epifluorescence and APC or PE immunofluorescence. The percentage of cells in each quadrant is indicated. Data shown are from a representative experiment performed three times with comparable results. Note preferential entry of HIV into CD4 cells and the absence of inhibitory effects of AMD3100 measured either at 3 or 24 h.

    Article Snippet: Cells were washed with phosphate-buffered saline (PBS) or treated with a trypsin-EDTA solution (1×; Sigma) for 2 min at room temperature to remove surface-bound virions, followed by additional washing and fixation in 1% paraformaldehyde-PBS solution.

    Techniques: Flow Cytometry, Infection, Labeling, Cell Culture, Cytometry, Derivative Assay, Incubation, Staining, Immunofluorescence

    Flow cytometric analysis of human SupT1 cells incubated with GFP-Vpr-labeled X4-tropic (A) or R5-tropic (B) HIV-1 virions in the presence of medium (panels 2 and 3), neutralizing anti-CD4 antibodies (panels 4 and 5), or AMD3100 (panels 6 and 7). SupT1 T cells were preincubated in the presence or absence of anti-CD4 antibodies or AMD3100 for 30 min at 37°C. Cells were then inoculated with GFP-Vpr-labeled X4-tropic HIV-1 or CCR5-tropic HIV-1 R5 (200 ng of p24 Gag) for 3 h at 37°C. The cells were subsequently washed with PBS (panels 1, 2, 4, and 6) or treated with a trypsin-EDTA solution (panels 3, 5, and 7) to remove surface-bound virions. Live cells were then analyzed by flow cytometry for GFP epifluorescence. The vertical bar indicates a gate established by analysis of uninfected cells (panels 1). The percentage of GFP-positive cells is indicated in the upper right hand corner of each panel. The anti-CD4 antibodies significantly inhibited the entry of both X4-tropic and R5-tropic virions, whereas AMD3100 unexpectedly failed to reduce the overall entry of X4-tropic virions.

    Journal: Journal of Virology

    Article Title: Compensatory Link between Fusion and Endocytosis of Human Immunodeficiency Virus Type 1 in Human CD4 T Lymphocytes

    doi: 10.1128/JVI.78.3.1375-1383.2004

    Figure Lengend Snippet: Flow cytometric analysis of human SupT1 cells incubated with GFP-Vpr-labeled X4-tropic (A) or R5-tropic (B) HIV-1 virions in the presence of medium (panels 2 and 3), neutralizing anti-CD4 antibodies (panels 4 and 5), or AMD3100 (panels 6 and 7). SupT1 T cells were preincubated in the presence or absence of anti-CD4 antibodies or AMD3100 for 30 min at 37°C. Cells were then inoculated with GFP-Vpr-labeled X4-tropic HIV-1 or CCR5-tropic HIV-1 R5 (200 ng of p24 Gag) for 3 h at 37°C. The cells were subsequently washed with PBS (panels 1, 2, 4, and 6) or treated with a trypsin-EDTA solution (panels 3, 5, and 7) to remove surface-bound virions. Live cells were then analyzed by flow cytometry for GFP epifluorescence. The vertical bar indicates a gate established by analysis of uninfected cells (panels 1). The percentage of GFP-positive cells is indicated in the upper right hand corner of each panel. The anti-CD4 antibodies significantly inhibited the entry of both X4-tropic and R5-tropic virions, whereas AMD3100 unexpectedly failed to reduce the overall entry of X4-tropic virions.

    Article Snippet: Cells were washed with phosphate-buffered saline (PBS) or treated with a trypsin-EDTA solution (1×; Sigma) for 2 min at room temperature to remove surface-bound virions, followed by additional washing and fixation in 1% paraformaldehyde-PBS solution.

    Techniques: Flow Cytometry, Incubation, Labeling, Cytometry