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Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a <t>FACS</t> machine with a <t>GFP</t> marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.
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1) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.
Figure Legend Snippet: Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.

Techniques Used: Transduction, Plasmid Preparation, Sequencing, FACS, Marker, Western Blot, Expressing

2) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Persistence of NK-92 cells in vivo . (a) Continuous persistence of NK-92 cells in the blood of mice by FACS. (b) The quantitative analysis results of NK-92 cells in tumors by FACS. (c) Representative results of NK-92 cells in tumors shown in (b). NK-92 cells were stained with APC-conjugated anti-hCD56 and APC-Cy7-conjugated anti-hCD45 antibodies in all experiments. ∗ p
Figure Legend Snippet: Persistence of NK-92 cells in vivo . (a) Continuous persistence of NK-92 cells in the blood of mice by FACS. (b) The quantitative analysis results of NK-92 cells in tumors by FACS. (c) Representative results of NK-92 cells in tumors shown in (b). NK-92 cells were stained with APC-conjugated anti-hCD56 and APC-Cy7-conjugated anti-hCD45 antibodies in all experiments. ∗ p

Techniques Used: In Vivo, Mouse Assay, FACS, Staining

3) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.
Figure Legend Snippet: Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.

Techniques Used: Transduction, Plasmid Preparation, Sequencing, FACS, Marker, Western Blot, Expressing

Specific cytokine release of EpCAM-specific CAR-NK-92 cells against EpCAM-positive cells. (a) FACS was used to test the surface expression of EpCAM proteins in 293T, 293T-EpCAM, FHC, and FHC-EpCAM cells and the human colorectal cancer cell lines HCT116, SW620, and HCT-8. (b) The levels of cytokines, released by Ctrl-NK-92 and CAR-NK-92 cells, were measured by enzyme-linked immunosorbent assay (ELISA) after incubation for 24 h with EpCAM-negative or EpCAM-positive target cells at an effector-to-target (E/T) ratio of 2 : 1. ∗∗ p
Figure Legend Snippet: Specific cytokine release of EpCAM-specific CAR-NK-92 cells against EpCAM-positive cells. (a) FACS was used to test the surface expression of EpCAM proteins in 293T, 293T-EpCAM, FHC, and FHC-EpCAM cells and the human colorectal cancer cell lines HCT116, SW620, and HCT-8. (b) The levels of cytokines, released by Ctrl-NK-92 and CAR-NK-92 cells, were measured by enzyme-linked immunosorbent assay (ELISA) after incubation for 24 h with EpCAM-negative or EpCAM-positive target cells at an effector-to-target (E/T) ratio of 2 : 1. ∗∗ p

Techniques Used: FACS, Expressing, Enzyme-linked Immunosorbent Assay, Incubation

4) Product Images from "Fluorescently Activated Cell Sorting Followed by Microarray Profiling of Helper T Cell Subtypes from Human Peripheral Blood"

Article Title: Fluorescently Activated Cell Sorting Followed by Microarray Profiling of Helper T Cell Subtypes from Human Peripheral Blood

Journal: PLoS ONE

doi: 10.1371/journal.pone.0111405

Schematic of the FACS–array procedure for peripheral blood cells. ( A ) Schematic of analysis of helper T (Th) cells using the FACS–array procedure for peripheral blood cells, where PBMCs are isolated on a density gradient centrifuge and stained with fluorescence-labeled antibodies (CD4-FITC, CXCR3-APC, and CCR4-PE) and PI for FACS. Total RNA was extracted from each subclass of lymphocytes and subjected to two rounds of cRNA amplification. Synthesized aminoallyl-aRNA samples were labeled with biotin and subjected to microarray analysis. ( B ) Dot plot imaging of FACS isolation of Th1 and Th2 cells. (1) Lymphocytic subpopulation of PBMCs was selected on the basis of their unique forward and side scatter properties on fluorocytometry. ( 2 ) Among the lymphocytes, PI-negative (viable) CD4 + Th cells were selected. (3) On the basis of the signal intensity of fluorostaining for the CXCR3 and CCR4 markers, the viable Th cells were separated into two subgroups: CXCR3 + /CCR4 − as Th1 cells and CXCR3 − /CCR4 + as Th2 cells.
Figure Legend Snippet: Schematic of the FACS–array procedure for peripheral blood cells. ( A ) Schematic of analysis of helper T (Th) cells using the FACS–array procedure for peripheral blood cells, where PBMCs are isolated on a density gradient centrifuge and stained with fluorescence-labeled antibodies (CD4-FITC, CXCR3-APC, and CCR4-PE) and PI for FACS. Total RNA was extracted from each subclass of lymphocytes and subjected to two rounds of cRNA amplification. Synthesized aminoallyl-aRNA samples were labeled with biotin and subjected to microarray analysis. ( B ) Dot plot imaging of FACS isolation of Th1 and Th2 cells. (1) Lymphocytic subpopulation of PBMCs was selected on the basis of their unique forward and side scatter properties on fluorocytometry. ( 2 ) Among the lymphocytes, PI-negative (viable) CD4 + Th cells were selected. (3) On the basis of the signal intensity of fluorostaining for the CXCR3 and CCR4 markers, the viable Th cells were separated into two subgroups: CXCR3 + /CCR4 − as Th1 cells and CXCR3 − /CCR4 + as Th2 cells.

Techniques Used: FACS, Isolation, Gradient Centrifugation, Staining, Fluorescence, Labeling, Amplification, Synthesized, Microarray, Imaging

5) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.
Figure Legend Snippet: Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.

Techniques Used: Transduction, Plasmid Preparation, Sequencing, FACS, Marker, Western Blot, Expressing

Specific cytokine release of EpCAM-specific CAR-NK-92 cells against EpCAM-positive cells. (a) FACS was used to test the surface expression of EpCAM proteins in 293T, 293T-EpCAM, FHC, and FHC-EpCAM cells and the human colorectal cancer cell lines HCT116, SW620, and HCT-8. (b) The levels of cytokines, released by Ctrl-NK-92 and CAR-NK-92 cells, were measured by enzyme-linked immunosorbent assay (ELISA) after incubation for 24 h with EpCAM-negative or EpCAM-positive target cells at an effector-to-target (E/T) ratio of 2 : 1. ∗∗ p
Figure Legend Snippet: Specific cytokine release of EpCAM-specific CAR-NK-92 cells against EpCAM-positive cells. (a) FACS was used to test the surface expression of EpCAM proteins in 293T, 293T-EpCAM, FHC, and FHC-EpCAM cells and the human colorectal cancer cell lines HCT116, SW620, and HCT-8. (b) The levels of cytokines, released by Ctrl-NK-92 and CAR-NK-92 cells, were measured by enzyme-linked immunosorbent assay (ELISA) after incubation for 24 h with EpCAM-negative or EpCAM-positive target cells at an effector-to-target (E/T) ratio of 2 : 1. ∗∗ p

Techniques Used: FACS, Expressing, Enzyme-linked Immunosorbent Assay, Incubation

6) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.
Figure Legend Snippet: Generation and characterization of EpCAM-specific CAR-NK-92 cells. (a) Structure diagram of EpCAM-specific CAR. EF1 α , promoter; SP, signal peptide; scFv, single-chain variable fragment. (b) Transduction efficiency of lentivirus in NK-92 cells. NK-92 cells were transduced with empty lentivirus vector (Ctrl-NK-92) or lentivirus containing the EpCAM-specific CAR encoding sequence (CAR-NK-92) and sorted by a FACS machine with a GFP marker. The GFP-positive rate of transduced NK-92 cells after sorting is shown. (c) Western blot analysis of the CAR expression in NK-92 cells with a monoclonal anti-human CD3 ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.

Techniques Used: Transduction, Plasmid Preparation, Sequencing, FACS, Marker, Western Blot, Expressing

7) Product Images from "Adventitial Progenitor Cells of Human Great Saphenous Vein Enhance the Resolution of Venous Thrombosis via Neovascularization"

Article Title: Adventitial Progenitor Cells of Human Great Saphenous Vein Enhance the Resolution of Venous Thrombosis via Neovascularization

Journal: Stem Cells International

doi: 10.1155/2021/8816763

Isolation and cultivation of CD34 + CD117 + HGSV-AdPC in vitro . (a) Cells of HGSV wall obtained by collagenase II digestion at 48 hours, 72 hours, and 7 days culture. The green arrow indicates cells growing in colony. (b) FACS isolation of CD34 + CD117 + HGSV-AdPC; the double positive cells were about (1.2 ± 0.07)% in vein wall cells. (c) Immunofluorescent staining with antibodies CD34 (red), CD117 (green), and DAPI (blue) of the FACS-purified HGSV-AdPC after 7 days culture, and the double-positive cells accounted for 91.2%. Scale bars: 200 μ m (a), 50 μ m (c). Magnification: 100x (a), 400x (c).
Figure Legend Snippet: Isolation and cultivation of CD34 + CD117 + HGSV-AdPC in vitro . (a) Cells of HGSV wall obtained by collagenase II digestion at 48 hours, 72 hours, and 7 days culture. The green arrow indicates cells growing in colony. (b) FACS isolation of CD34 + CD117 + HGSV-AdPC; the double positive cells were about (1.2 ± 0.07)% in vein wall cells. (c) Immunofluorescent staining with antibodies CD34 (red), CD117 (green), and DAPI (blue) of the FACS-purified HGSV-AdPC after 7 days culture, and the double-positive cells accounted for 91.2%. Scale bars: 200 μ m (a), 50 μ m (c). Magnification: 100x (a), 400x (c).

Techniques Used: Isolation, In Vitro, FACS, Staining, Purification

CD34 + CD117 + HGSV-AdPCs differentiated into endothelial cells in vitro . CD34 + CD117 + HGSV-AdPCs were isolated with FACS and cultured in the presence of VEGF for 7 days. (a) The mRNA expression of CD31, VEGFR2, and vWF. (b) The result of Western blotting of CD31 and VEGFR2 proteins in the induction and control groups. (c) The differentiation result of immunofluorescent staining with antibodies CD31 and VEGFR2 in control and induction groups. (d) The tube formation observed under inverse microscope at 6 hours culture in control and induction groups. (e) The difference of tube formation at 6 hours culture between control and induction groups. Scale bars: 10 μ m (c); 200 μ m (d). Magnification: 200x (a, d).
Figure Legend Snippet: CD34 + CD117 + HGSV-AdPCs differentiated into endothelial cells in vitro . CD34 + CD117 + HGSV-AdPCs were isolated with FACS and cultured in the presence of VEGF for 7 days. (a) The mRNA expression of CD31, VEGFR2, and vWF. (b) The result of Western blotting of CD31 and VEGFR2 proteins in the induction and control groups. (c) The differentiation result of immunofluorescent staining with antibodies CD31 and VEGFR2 in control and induction groups. (d) The tube formation observed under inverse microscope at 6 hours culture in control and induction groups. (e) The difference of tube formation at 6 hours culture between control and induction groups. Scale bars: 10 μ m (c); 200 μ m (d). Magnification: 200x (a, d).

Techniques Used: In Vitro, Isolation, FACS, Cell Culture, Expressing, Western Blot, Staining, Microscopy

8) Product Images from "Reactivation of Lytic Replication from B Cells Latently Infected with Epstein-Barr Virus Occurs with High S-Phase Cyclin-Dependent Kinase Activity while Inhibiting Cellular DNA Replication"

Article Title: Reactivation of Lytic Replication from B Cells Latently Infected with Epstein-Barr Virus Occurs with High S-Phase Cyclin-Dependent Kinase Activity while Inhibiting Cellular DNA Replication

Journal: Journal of Virology

doi: 10.1128/JVI.77.2.851-861.2003

Lytic program arrests cell cycle progression of Tet-BZLF1/B95-8 cells. Tet-BZLF1/B95-8 cells were cultured in the presence [Dox (+)] or absence [Dox (−)] of 1 μg of doxycycline/ml for 72 h. If necessary, paclitaxel (20 μM) was added at 48 h p.i. to arrest the cell cycle at M phase, and the cells were incubated for another 24 h (top). After fixation with methanol, the cells were doubly stained with anti-BMRF1 IgG and PI and analyzed by FACS. The cell cycle distribution was analyzed in the total cell population [Dox (−)] and the BMRF1-positive population [Dox (+)] by using ModFit LT software (Becton Dickinson).
Figure Legend Snippet: Lytic program arrests cell cycle progression of Tet-BZLF1/B95-8 cells. Tet-BZLF1/B95-8 cells were cultured in the presence [Dox (+)] or absence [Dox (−)] of 1 μg of doxycycline/ml for 72 h. If necessary, paclitaxel (20 μM) was added at 48 h p.i. to arrest the cell cycle at M phase, and the cells were incubated for another 24 h (top). After fixation with methanol, the cells were doubly stained with anti-BMRF1 IgG and PI and analyzed by FACS. The cell cycle distribution was analyzed in the total cell population [Dox (−)] and the BMRF1-positive population [Dox (+)] by using ModFit LT software (Becton Dickinson).

Techniques Used: Cell Culture, Incubation, Staining, FACS, Software

9) Product Images from "Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models"

Article Title: Combination Therapy with EpCAM-CAR-NK-92 Cells and Regorafenib against Human Colorectal Cancer Models

Journal: Journal of Immunology Research

doi: 10.1155/2018/4263520

Persistence of NK-92 cells in vivo . (a) Continuous persistence of NK-92 cells in the blood of mice by FACS. (b) The quantitative analysis results of NK-92 cells in tumors by FACS. (c) Representative results of NK-92 cells in tumors shown in (b). NK-92 cells were stained with APC-conjugated anti-hCD56 and APC-Cy7-conjugated anti-hCD45 antibodies in all experiments. ∗ p
Figure Legend Snippet: Persistence of NK-92 cells in vivo . (a) Continuous persistence of NK-92 cells in the blood of mice by FACS. (b) The quantitative analysis results of NK-92 cells in tumors by FACS. (c) Representative results of NK-92 cells in tumors shown in (b). NK-92 cells were stained with APC-conjugated anti-hCD56 and APC-Cy7-conjugated anti-hCD45 antibodies in all experiments. ∗ p

Techniques Used: In Vivo, Mouse Assay, FACS, Staining

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Mouse Assay:

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Staining:

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Flow Cytometry:

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Labeling:

Article Title: Single-walled carbon-nanohorns improve biocompatibility over nanotubes by triggering less protein-initiated pyroptosis and apoptosis in macrophages
Article Snippet: The obtained cell suspensions were added with Annexin V-FITC-binding solution for 30 min at room temperature and further incubated with PI staining solution for 5 min before detection. .. The labeled cells were then analyzed by flow cytometry system (FACS Calibur, BD, NJ, USA). .. Data were treated by FlowJo 7.6 software.

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Incubation:

Article Title: Single-walled carbon-nanohorns improve biocompatibility over nanotubes by triggering less protein-initiated pyroptosis and apoptosis in macrophages
Article Snippet: The difference of cell deaths caused by five nanocarbons was evaluated by flow cytometry based on PI labeling. .. In the investigation, cells were incubated with nanocarbons (100 μg ml−1 ) at 37 °C for 24 h. After washing with PBS, cells were performed by cell scraper to capture single-cell suspensions, labeled by PI solution (2 μg ml−1) at 4 °C for 20 min, and finally detected by flow cytometry system (FACS Calibur, BD, NJ, USA) at the PI channel. .. The data were performed by FlowJo 7.6 software.

Article Title: GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
Article Snippet: .. Following 3-day incubation at 37°C, NK cells were collected and assessed by Fluorescence Activated Cell Sorting (FACS) analysis with APC-conjugated anti-CD45, BV786-conjugated anti-CD16, BV711-conjugated anti-CD25, BUV395-conjiugated anti-CD56 (BD Biosciences, CA-USA) and PE-conjugated anti-NKp44, PE-Cy7-conjugated anti-CD69 antibodies (Immunological Science, SIC, IT). .. For apoptosis evaluation, target tumor cells were harvested at the indicated time points and incubated with APC-conjugated anti-CD45 and PE-conjugated anti-Annexin V antibodies following the manufacturer-s instructions (Immunological Sciences, SIC, IT).

Article Title: TMEM41B Is a Pan-flavivirus Host Factor
Article Snippet: For non-fluorescent reporter viruses, cells were washed 1x in PBS (+ 2% FBS), permeabilized in 1x BD perm (BD Biosciences: 554723) for 1 h at RT followed by overnight incubation with primary antibody. .. Cells were then washed three times with BD perm, incubated with secondary antibody for 1 h at RT, washed an additional three times with BD perm, and once with PBS (+ 2% FBS) before being transferred to FACS tubes for flow-cytometry analysis similar as for aforementioned reporter viruses. .. Cells infected in 96-well plates were fixed with 10% neutral-buffered formalin, permeabilized with 0.1% Triton X-100 in PBS for 10 min at RT, washed in PBS with 0.01% Tween20 (PBS-T) and incubated with primary antibody (pan-flavivirus HMAF) overnight.

Fluorescence:

Article Title: GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
Article Snippet: .. Following 3-day incubation at 37°C, NK cells were collected and assessed by Fluorescence Activated Cell Sorting (FACS) analysis with APC-conjugated anti-CD45, BV786-conjugated anti-CD16, BV711-conjugated anti-CD25, BUV395-conjiugated anti-CD56 (BD Biosciences, CA-USA) and PE-conjugated anti-NKp44, PE-Cy7-conjugated anti-CD69 antibodies (Immunological Science, SIC, IT). .. For apoptosis evaluation, target tumor cells were harvested at the indicated time points and incubated with APC-conjugated anti-CD45 and PE-conjugated anti-Annexin V antibodies following the manufacturer-s instructions (Immunological Sciences, SIC, IT).

Expressing:

Article Title: Regulation of DNA Replication Timing on Human Chromosome by a Cell-Type Specific DNA Binding Protein SATB1
Article Snippet: Cells were collected in PBS containing of 2 µg/ml propidium iodide (SIGMA). .. Living cells expressing mKO2 were sorted by FACS Aria (BD Biosciences). .. The sorted mKO2-positive cells were used for replication timing analysis by FISH as described above.

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    Becton Dickinson facs aria iii
    Self-reactive CD4 + T cells capable of inducing autoimmune response accumulate in the periphery in the absence of B7-CD28 co-stimulation. a Peripheral pMOG-specific CD4 + T cell number was increased in B7 DKO mice. Each group n = 5. Data shown are combined results of at least <t>three</t> independent experiments. Tconv p = 0.0002, Treg p = 0.0114. Data are mean ± SEM with dots representing individual values of biologically independent animals. Statistical differences between groups were calculated using unpaired, two-tailed Student’s t -test. b pMOG-specific peripheral CD4 + T cells do not have an anergic phenotype. Each group n = 4. Data are a representative <t>FACS</t> plot of three independent experiments. c CD4 + T cells from B7 DKO mice strongly induced EAE. WT n = 12, B7 DKO n = 13 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0089. Medians of the total clinical score during day 25–37 were compared by two-tailed non-parametric Mann–Whitney test. d Treg-depleted CD4 + T conv cells from B7 DKO mice strongly induced EAE. WT n = 10, B7 DKO n = 10 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0011. Medians of the total clinical score during day 15–35 were compared by two-tailed non-parametric Mann–Whitney test. * p
    Facs Aria Iii, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson facs canto system
    Effect of enhanced expression of NAG-1 on TA-induced <t>apoptosis</t> in KB cells. KB/NAG-1 and KB/vector cells were treated with 30 µM TA for 24 h, and apoptosis was analyzed using a TUNEL assay and by <t>FACS</t> with annexin V-FITC/PI. (A) Western Blot. Equal amounts of cell lysates (30 µg) were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane, and probed with anti-NAG-1, PARP, cleaved caspase-3, or α–tubulin antibody. (B and C) Apoptosis in KB cells was determined by the TUNEL method and FACS with AnnexinV-FITC/PI, as described in Fig. 3 . Data are mean values obtained from three independent experiments and bars represent standard deviations. The scale bar denotes 50 µm.
    Facs Canto System, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson facs canto ii flow cytometer
    Induction of GIL-specific T cell responses by different concentrations of GIL and whole-inactivated influenza virus (WIV). HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with indicated amounts of GIL peptide and WIV. (A) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine <t>IFN-γ.</t> The percentages for IFN-γ + CD8 + T cells acquired by <t>FACS</t> are plotted. (B) Splenocytes of immunized mice were re-stimulated with GIL peptide. The number of CD8 + IFN-γ-secreting cells was subsequently determined with ELISpot. (A,B) Data are shown as mean ± SD of three mice per group, each dot is a single replicate and data are from a single experiment. * p
    Facs Canto Ii Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson facs analysis
    Enforced expression of AID rescues CSR in Hoxc4 −/− B cells. Hoxc4 +/+ and Hoxc4 −/− B cells activated with LPS were transduced with the TAC control or AID-expression TAC- Aicda retrovirus and cultured in the presence of LPS and IL-4. Three or 4 d after transduction, B cells were harvested for analysis of surface expression of B220 and <t>IgG1</t> ( a ) and expression of Aicda by real-time qRT-PCR and semi-quantitative RT-PCR, germline I µ -C µ and I γ 1-C γ 1 transcripts by real-time qRT-PCR, circle I γ 1-C µ transcripts by semi-quantitative RT-PCR and post-recombination I µ -C γ 1 transcripts by real-time qRT-PCR ( b ). Expression of these transcripts was normalized in each case to CD79b transcripts; expression of transcripts in Hoxc4 −/− B cells were depicted as ratios to those in Hoxc4 +/+ B cells. <t>FACS</t> data are from one representative of 5 independent experiments. Real-time qRT-PCR and semi-quantitative RT-PCR data are means ± s.e. (bars) of 3 independent experiments.
    Facs Analysis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Self-reactive CD4 + T cells capable of inducing autoimmune response accumulate in the periphery in the absence of B7-CD28 co-stimulation. a Peripheral pMOG-specific CD4 + T cell number was increased in B7 DKO mice. Each group n = 5. Data shown are combined results of at least three independent experiments. Tconv p = 0.0002, Treg p = 0.0114. Data are mean ± SEM with dots representing individual values of biologically independent animals. Statistical differences between groups were calculated using unpaired, two-tailed Student’s t -test. b pMOG-specific peripheral CD4 + T cells do not have an anergic phenotype. Each group n = 4. Data are a representative FACS plot of three independent experiments. c CD4 + T cells from B7 DKO mice strongly induced EAE. WT n = 12, B7 DKO n = 13 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0089. Medians of the total clinical score during day 25–37 were compared by two-tailed non-parametric Mann–Whitney test. d Treg-depleted CD4 + T conv cells from B7 DKO mice strongly induced EAE. WT n = 10, B7 DKO n = 10 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0011. Medians of the total clinical score during day 15–35 were compared by two-tailed non-parametric Mann–Whitney test. * p

    Journal: Nature Communications

    Article Title: B7-CD28 co-stimulation modulates central tolerance via thymic clonal deletion and Treg generation through distinct mechanisms

    doi: 10.1038/s41467-020-20070-x

    Figure Lengend Snippet: Self-reactive CD4 + T cells capable of inducing autoimmune response accumulate in the periphery in the absence of B7-CD28 co-stimulation. a Peripheral pMOG-specific CD4 + T cell number was increased in B7 DKO mice. Each group n = 5. Data shown are combined results of at least three independent experiments. Tconv p = 0.0002, Treg p = 0.0114. Data are mean ± SEM with dots representing individual values of biologically independent animals. Statistical differences between groups were calculated using unpaired, two-tailed Student’s t -test. b pMOG-specific peripheral CD4 + T cells do not have an anergic phenotype. Each group n = 4. Data are a representative FACS plot of three independent experiments. c CD4 + T cells from B7 DKO mice strongly induced EAE. WT n = 12, B7 DKO n = 13 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0089. Medians of the total clinical score during day 25–37 were compared by two-tailed non-parametric Mann–Whitney test. d Treg-depleted CD4 + T conv cells from B7 DKO mice strongly induced EAE. WT n = 10, B7 DKO n = 10 biologically independent animals. Data are pooled results of two independent experiments (mean ± SEM). p = 0.0011. Medians of the total clinical score during day 15–35 were compared by two-tailed non-parametric Mann–Whitney test. * p

    Article Snippet: Data were collected with a FACS Calibur II, FACS LSR II, FACS Fortessa, or FACS Aria III (BD Biosciences) flow cytometer and analyzed with FlowJo software (Tree Star).

    Techniques: Mouse Assay, Two Tailed Test, FACS, MANN-WHITNEY

    Effect of enhanced expression of NAG-1 on TA-induced apoptosis in KB cells. KB/NAG-1 and KB/vector cells were treated with 30 µM TA for 24 h, and apoptosis was analyzed using a TUNEL assay and by FACS with annexin V-FITC/PI. (A) Western Blot. Equal amounts of cell lysates (30 µg) were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane, and probed with anti-NAG-1, PARP, cleaved caspase-3, or α–tubulin antibody. (B and C) Apoptosis in KB cells was determined by the TUNEL method and FACS with AnnexinV-FITC/PI, as described in Fig. 3 . Data are mean values obtained from three independent experiments and bars represent standard deviations. The scale bar denotes 50 µm.

    Journal: PLoS ONE

    Article Title: Tolfenamic Acid Induces Apoptosis and Growth Inhibition in Head and Neck Cancer: Involvement of NAG-1 Expression

    doi: 10.1371/journal.pone.0034988

    Figure Lengend Snippet: Effect of enhanced expression of NAG-1 on TA-induced apoptosis in KB cells. KB/NAG-1 and KB/vector cells were treated with 30 µM TA for 24 h, and apoptosis was analyzed using a TUNEL assay and by FACS with annexin V-FITC/PI. (A) Western Blot. Equal amounts of cell lysates (30 µg) were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane, and probed with anti-NAG-1, PARP, cleaved caspase-3, or α–tubulin antibody. (B and C) Apoptosis in KB cells was determined by the TUNEL method and FACS with AnnexinV-FITC/PI, as described in Fig. 3 . Data are mean values obtained from three independent experiments and bars represent standard deviations. The scale bar denotes 50 µm.

    Article Snippet: Apoptosis was detected using a FACS Canto system (BD Biosciences, Bedford, MA), with excitation and emission settings of 488 and 530 nm, respectively.

    Techniques: Expressing, Plasmid Preparation, TUNEL Assay, FACS, Western Blot, Polyacrylamide Gel Electrophoresis, SDS Page

    Effect of suppression of NAG-1 expression on TA-induced apoptosis in KB cells. KB cells transfected with either the indicated NAG-1 siRNA or negative control siRNA were treated with or without 30 µM TA for 24 h. Apoptosis was analyzed by the TUNEL assay and by FACS with AnnexinV-FITC/PI. (A) Western Blot. Equal amounts of cell lysates (30 µg) were resolved by SDS-PAGE, transferred to nitrocellulose membrane, and probed with anti- NAG-1, PARP, cleaved caspase-3, orαα–tubulin. (B and C) Apoptosis in KB cells was determined by the TUNEL method and FACS with AnnexinV-FITC/PI, as described in Fig. 3 . Data are mean values obtained from three independent experiments and bars represent standard deviations. The scale bar denotes 50 µm.

    Journal: PLoS ONE

    Article Title: Tolfenamic Acid Induces Apoptosis and Growth Inhibition in Head and Neck Cancer: Involvement of NAG-1 Expression

    doi: 10.1371/journal.pone.0034988

    Figure Lengend Snippet: Effect of suppression of NAG-1 expression on TA-induced apoptosis in KB cells. KB cells transfected with either the indicated NAG-1 siRNA or negative control siRNA were treated with or without 30 µM TA for 24 h. Apoptosis was analyzed by the TUNEL assay and by FACS with AnnexinV-FITC/PI. (A) Western Blot. Equal amounts of cell lysates (30 µg) were resolved by SDS-PAGE, transferred to nitrocellulose membrane, and probed with anti- NAG-1, PARP, cleaved caspase-3, orαα–tubulin. (B and C) Apoptosis in KB cells was determined by the TUNEL method and FACS with AnnexinV-FITC/PI, as described in Fig. 3 . Data are mean values obtained from three independent experiments and bars represent standard deviations. The scale bar denotes 50 µm.

    Article Snippet: Apoptosis was detected using a FACS Canto system (BD Biosciences, Bedford, MA), with excitation and emission settings of 488 and 530 nm, respectively.

    Techniques: Expressing, Transfection, Negative Control, TUNEL Assay, FACS, Western Blot, SDS Page

    Induction of GIL-specific T cell responses by different concentrations of GIL and whole-inactivated influenza virus (WIV). HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with indicated amounts of GIL peptide and WIV. (A) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (B) Splenocytes of immunized mice were re-stimulated with GIL peptide. The number of CD8 + IFN-γ-secreting cells was subsequently determined with ELISpot. (A,B) Data are shown as mean ± SD of three mice per group, each dot is a single replicate and data are from a single experiment. * p

    Journal: Frontiers in Immunology

    Article Title: Whole-Inactivated Influenza Virus Is a Potent Adjuvant for Influenza Peptides Containing CD8+ T Cell Epitopes

    doi: 10.3389/fimmu.2018.00525

    Figure Lengend Snippet: Induction of GIL-specific T cell responses by different concentrations of GIL and whole-inactivated influenza virus (WIV). HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with indicated amounts of GIL peptide and WIV. (A) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (B) Splenocytes of immunized mice were re-stimulated with GIL peptide. The number of CD8 + IFN-γ-secreting cells was subsequently determined with ELISpot. (A,B) Data are shown as mean ± SD of three mice per group, each dot is a single replicate and data are from a single experiment. * p

    Article Snippet: Finally, cells were stained intracellularly with anti-mouse IFN-γ-APC (BD Biosciences), and IFN-γ+ CD8+ T cells were quantified on a FACS Canto II flow cytometer (BD Biosciences).

    Techniques: Transgenic Assay, Mouse Assay, Staining, FACS, Enzyme-linked Immunospot

    Comparison of CD8 + T cell responses induced by 1 nmol GIL peptide adjuvanted with either 50 µg CpG or 50 µg whole-inactivated influenza virus (WIV). HLA-A2.1 transgenic mice were injected twice, 3 weeks apart with PBS (negative control), WIV or GIL peptide with indicated adjuvant. Shown are the responses 2 weeks after the final immunization. (A) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. Data are shown as mean ± SD of three mice per group, each circle is a single replicate and data are from a single experiment representative of two individual experiments. ** p

    Journal: Frontiers in Immunology

    Article Title: Whole-Inactivated Influenza Virus Is a Potent Adjuvant for Influenza Peptides Containing CD8+ T Cell Epitopes

    doi: 10.3389/fimmu.2018.00525

    Figure Lengend Snippet: Comparison of CD8 + T cell responses induced by 1 nmol GIL peptide adjuvanted with either 50 µg CpG or 50 µg whole-inactivated influenza virus (WIV). HLA-A2.1 transgenic mice were injected twice, 3 weeks apart with PBS (negative control), WIV or GIL peptide with indicated adjuvant. Shown are the responses 2 weeks after the final immunization. (A) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. Data are shown as mean ± SD of three mice per group, each circle is a single replicate and data are from a single experiment representative of two individual experiments. ** p

    Article Snippet: Finally, cells were stained intracellularly with anti-mouse IFN-γ-APC (BD Biosciences), and IFN-γ+ CD8+ T cells were quantified on a FACS Canto II flow cytometer (BD Biosciences).

    Techniques: Transgenic Assay, Mouse Assay, Injection, Negative Control, Staining, FACS

    Influence of co-localization on the immunogenicity of GIL (100 nmol) and whole-inactivated influenza virus (WIV, 25 µg). (A) HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with GIL peptide and WIV either combined in one single flank (mixed) or separate in opposite flanks (separate). (B) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (C) Splenocytes were re-stimulated with GIL peptide. The number of IFN-γ-secreting cells was subsequently determined with ELISpot. (A,B) Data are shown as mean ± SD of six mice per group, each dot is a single replicate and data are from a single experiment. ** p

    Journal: Frontiers in Immunology

    Article Title: Whole-Inactivated Influenza Virus Is a Potent Adjuvant for Influenza Peptides Containing CD8+ T Cell Epitopes

    doi: 10.3389/fimmu.2018.00525

    Figure Lengend Snippet: Influence of co-localization on the immunogenicity of GIL (100 nmol) and whole-inactivated influenza virus (WIV, 25 µg). (A) HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with GIL peptide and WIV either combined in one single flank (mixed) or separate in opposite flanks (separate). (B) Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (C) Splenocytes were re-stimulated with GIL peptide. The number of IFN-γ-secreting cells was subsequently determined with ELISpot. (A,B) Data are shown as mean ± SD of six mice per group, each dot is a single replicate and data are from a single experiment. ** p

    Article Snippet: Finally, cells were stained intracellularly with anti-mouse IFN-γ-APC (BD Biosciences), and IFN-γ+ CD8+ T cells were quantified on a FACS Canto II flow cytometer (BD Biosciences).

    Techniques: Transgenic Assay, Mouse Assay, Staining, FACS, Enzyme-linked Immunospot

    Effect of membrane fusion activity on whole-inactivated influenza virus (WIV) adjuvant activity. (A) WIV was fusion-inactivated by brief exposure to a buffer with pH 4.5 at 37°C. Hemolysis was performed by mixing either WIV (“active” WIV) or fusion-inactivated WIV (“inactive” WIV) with human blood erythrocytes at various pHs. Fusion-mediated hemoglobin release was subsequently determined by spectrophotometric analysis. Data are shown as mean ± SD of three individual experiments. (B) HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with GIL peptide (100 µg) and either fusion-active or fusion-inactive WIV (25 µg). Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (C) Splenocytes were re-stimulated with GIL peptide. The number of IFN-γ-secreting cells was subsequently determined with ELISpot. (B,C) Data are shown as mean ± SD of six mice per group, each dot is a single replicate and data are from a single experiment. n.s. = not significant (one-way ANOVA).

    Journal: Frontiers in Immunology

    Article Title: Whole-Inactivated Influenza Virus Is a Potent Adjuvant for Influenza Peptides Containing CD8+ T Cell Epitopes

    doi: 10.3389/fimmu.2018.00525

    Figure Lengend Snippet: Effect of membrane fusion activity on whole-inactivated influenza virus (WIV) adjuvant activity. (A) WIV was fusion-inactivated by brief exposure to a buffer with pH 4.5 at 37°C. Hemolysis was performed by mixing either WIV (“active” WIV) or fusion-inactivated WIV (“inactive” WIV) with human blood erythrocytes at various pHs. Fusion-mediated hemoglobin release was subsequently determined by spectrophotometric analysis. Data are shown as mean ± SD of three individual experiments. (B) HLA-A2.1 transgenic mice were vaccinated twice, 3 weeks apart with GIL peptide (100 µg) and either fusion-active or fusion-inactive WIV (25 µg). Splenocytes were re-stimulated with GIL peptide and stained for intracellular cytokine IFN-γ. The percentages for IFN-γ + CD8 + T cells acquired by FACS are plotted. (C) Splenocytes were re-stimulated with GIL peptide. The number of IFN-γ-secreting cells was subsequently determined with ELISpot. (B,C) Data are shown as mean ± SD of six mice per group, each dot is a single replicate and data are from a single experiment. n.s. = not significant (one-way ANOVA).

    Article Snippet: Finally, cells were stained intracellularly with anti-mouse IFN-γ-APC (BD Biosciences), and IFN-γ+ CD8+ T cells were quantified on a FACS Canto II flow cytometer (BD Biosciences).

    Techniques: Activity Assay, Transgenic Assay, Mouse Assay, Staining, FACS, Enzyme-linked Immunospot

    Enforced expression of AID rescues CSR in Hoxc4 −/− B cells. Hoxc4 +/+ and Hoxc4 −/− B cells activated with LPS were transduced with the TAC control or AID-expression TAC- Aicda retrovirus and cultured in the presence of LPS and IL-4. Three or 4 d after transduction, B cells were harvested for analysis of surface expression of B220 and IgG1 ( a ) and expression of Aicda by real-time qRT-PCR and semi-quantitative RT-PCR, germline I µ -C µ and I γ 1-C γ 1 transcripts by real-time qRT-PCR, circle I γ 1-C µ transcripts by semi-quantitative RT-PCR and post-recombination I µ -C γ 1 transcripts by real-time qRT-PCR ( b ). Expression of these transcripts was normalized in each case to CD79b transcripts; expression of transcripts in Hoxc4 −/− B cells were depicted as ratios to those in Hoxc4 +/+ B cells. FACS data are from one representative of 5 independent experiments. Real-time qRT-PCR and semi-quantitative RT-PCR data are means ± s.e. (bars) of 3 independent experiments.

    Journal: Nature immunology

    Article Title: HoxC4 binds to the Aicda promoter to induce AID expression, class switch DNA recombination and somatic hypermutation

    doi: 10.1038/ni.1725

    Figure Lengend Snippet: Enforced expression of AID rescues CSR in Hoxc4 −/− B cells. Hoxc4 +/+ and Hoxc4 −/− B cells activated with LPS were transduced with the TAC control or AID-expression TAC- Aicda retrovirus and cultured in the presence of LPS and IL-4. Three or 4 d after transduction, B cells were harvested for analysis of surface expression of B220 and IgG1 ( a ) and expression of Aicda by real-time qRT-PCR and semi-quantitative RT-PCR, germline I µ -C µ and I γ 1-C γ 1 transcripts by real-time qRT-PCR, circle I γ 1-C µ transcripts by semi-quantitative RT-PCR and post-recombination I µ -C γ 1 transcripts by real-time qRT-PCR ( b ). Expression of these transcripts was normalized in each case to CD79b transcripts; expression of transcripts in Hoxc4 −/− B cells were depicted as ratios to those in Hoxc4 +/+ B cells. FACS data are from one representative of 5 independent experiments. Real-time qRT-PCR and semi-quantitative RT-PCR data are means ± s.e. (bars) of 3 independent experiments.

    Article Snippet: B and T cells The number of B cells (B220+ ) and T cells (CD3+ ), the proportion CD4+ T cells, dead B and T cells and the proportion of PNAhi B cells, plasma cells (B220lo CD138+ ) and NP-binding CD38hi IgG1+ memory B cells were determined by FACS analysis using a FACSCalibur ™ flow cytometer (BD Biosciences).

    Techniques: Expressing, Transduction, Cell Culture, Quantitative RT-PCR, FACS