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Becton Dickinson flow cytometry
Flow Cytometry, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 36223 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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flow cytometry - by Bioz Stars, 2020-05
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Related Articles

Flow Cytometry:

Article Title: Selection of antitumor displayed peptides for the specific delivery of the anticancer drug lactaptin
Article Snippet: .. The exposure of phosphatidylserine (PS) at the outer surface of the cell membrane, and the activation of caspase-3 and −7 were analyzed by flow cytofluorometry with fluorescein isothiocyanate Annexin V Apoptosis Detection kit I (BD Pharmingen, San Jose, CA, USA) and the Vybrant FAM Caspase-3 and −7 assay kit (Life Technologies; Thermo Fisher Scientific, Inc.) using a FACSCanto II cell analyzer (BD Biosciences, Franklin Lakes, NJ, USA). .. Labeling of the recombinant lactaptin analogs with sulfo-Cy5 The recombinant lactaptin analogs RL2, Т1_RL and Т2_RL were isolated from E. coli and purified according to a protocol for the isolation of RL2 ( ).

Article Title: Estrogenic Activity of Coumestrol, DDT, and TCDD in Human Cervical Cancer Cells
Article Snippet: .. Flow microfluorometry was performed and DNA histograms were generated and analyzed using a Becton Dickinson Flowscan (Franklin Lakes, NJ). .. This method correlates closely with other measures of apoptosis including TUNEL and Annexin V staining while providing additional cell cycle information [ ].

Article Title: Identification of a Boundary Domain Adjacent to the Potent Human Cytomegalovirus Enhancer That Represses Transcription of the Divergent UL127 Promoter †
Article Snippet: .. Fluorescence was measured by flow cytofluorometry (fluorescence-activated cell sorting [FACS]) using a FACScan (Becton Dickinson, Mountain View, Calif.). .. Uncompensated fluorescence values from FL1 (GFP fluorescence, 530-nm band pass filter) and FL2 (autofluorescence; a 585-nm band pass filter) were collected along with forward angle light scatter (FSC) and side scatter (SSC) for every event.

Article Title: Effect of CHK1 Inhibition on CPX-351 Cytotoxicity in vitro and ex vivo
Article Snippet: .. Analysis of cell cycle distribution and apoptosis After incubation for 24 h with CPX-351 in the absence or presence of CHK1 inhibitors, cells were resuspended in ice cold 0.1% (wt/vol) sodium citrate containing 50 µg/mL PI and 0.1% (wt/vol) Triton X-100, incubated at 4 °C overnight, and analyzed by flow microfluorimetry in the FL2 channel on a Becton Dickinson (Franklin Lakes, NJ) FASCanto II flow cytometer. .. After collection of 20,000 events, files were analyzed using Modfit (Verity Software, Topsham, ME) for cell cycle distribution or Becton Dickinson CellQuest software for events with < 2n DNA content.

Article Title: IgE-FcεRI Interactions Determine HIV Coreceptor Usage and Susceptibility to Infection during Ontogeny of Mast Cells 1
Article Snippet: .. Analysis of the surface expression of CXCR4 and Fc ε RI-bound IgE on paraformaldehyde-treated prMC was performed by immunophenotyping and flow microfluorometry with a BD Biosciences FACSCalibur using CellQuest software as reported previously ( ). ..

Article Title: Aberrant populations of circulating T follicular helper cells and regulatory B cells underlying idiopathic pulmonary fibrosis
Article Snippet: .. We isolated heparinized peripheral blood mononuclear cells from fresh blood specimens by centrifugation over a discontinuous density gradient (Lympholyte-H; Cedarlane Laboratories Ltd., Canada) and investigated the proportion of Breg cells (to CD3− CD19+ B cells), Tfh cells (to CD3+ CD4+ T cells), PD-1+ ICOS+ Tfh cells (to all Tfh cells), and the Tfh-cell subset by flow cytometry (BD FACSCanto™ II; BD Biosciences, USA). ..

Article Title: NF-κB, p38 MAPK, ERK1/2, mTOR, STAT3 and increased glycolysis regulate stability of paricalcitol/dexamethasone-generated tolerogenic dendritic cells in the inflammatory environment
Article Snippet: .. Reagents and Abs Flow cytometry: commercial antibodies anti-CD86-FITC (clone 2231 FUN-1), CD274 (PD-L1)-FITC (clone MIH1), CD273 (PD-L2)-PE (clone MIH-18), HLA-DR-PE-Cy7 (clone L243), IFN-γ-FITC (clone 4SB3) were purchased from BD Biosciences; CD83-PerCP-Cy5.5 (clone HB15a) was purchased from Beckman Coulter; CD80-FITC (clone MAB104), CD40-PerCP-eFluor710 (clone 5C3), CD1a-PE-Cy7 (clone HI149) and CD4-PE-Cy7 (clone RPA-T4) were purchased from eBioscience; TLR2-FITC (clone T2.5), TIM-3-PE (clone F38-2E2), IL-10-PE (clone JES3-9D7), KI-67-PE (clone Ki-67) were purchased from BioLegend; CD14-PE-DL594 (clone MEM-15), CD11c-APC (clone BU15), CD3-AF700 (clone MEM-57), CD8-PE-Dy590 (clone MEM-31) were purchased from Exbio; CD85k (ILT-3)-PE (clone 293623), CD85d (IL-T4)-FITC (clone 287219) were purchased from R & D Systems. .. For western blot, anti-p-p38 MAPK, anti-p-ERK1/2, anti-p-JNK/SAPK, anti-p-IκB-α, anti-IDO, anti-p-mTOR, anti-p-STAT3, anti-p-p70S6K, anti-p38 MAPK, anti-ERK1/2, anti-JNK/SAPK and anti-STAT3 Ab were purchased from Cell Signaling Technology; anti-actin was from BioLegend.

Article Title: Absence of phagocyte NADPH oxidase 2 leads to severe inflammatory response in lungs of mice infected with Coccidioides
Article Snippet: .. In brief, absolute numbers of pulmonary leukocytes in the lung tissue preparations were enumerated by flow cytofluorometry using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ). .. Data were acquired with CellQuest Pro software (BD Biosciences) and analyzed using a FlowJo software package (Tree Star, Inc., Ashland, OR).

Fluorescence:

Article Title: Identification of a Boundary Domain Adjacent to the Potent Human Cytomegalovirus Enhancer That Represses Transcription of the Divergent UL127 Promoter †
Article Snippet: .. Fluorescence was measured by flow cytofluorometry (fluorescence-activated cell sorting [FACS]) using a FACScan (Becton Dickinson, Mountain View, Calif.). .. Uncompensated fluorescence values from FL1 (GFP fluorescence, 530-nm band pass filter) and FL2 (autofluorescence; a 585-nm band pass filter) were collected along with forward angle light scatter (FSC) and side scatter (SSC) for every event.

Isolation:

Article Title: Aberrant populations of circulating T follicular helper cells and regulatory B cells underlying idiopathic pulmonary fibrosis
Article Snippet: .. We isolated heparinized peripheral blood mononuclear cells from fresh blood specimens by centrifugation over a discontinuous density gradient (Lympholyte-H; Cedarlane Laboratories Ltd., Canada) and investigated the proportion of Breg cells (to CD3− CD19+ B cells), Tfh cells (to CD3+ CD4+ T cells), PD-1+ ICOS+ Tfh cells (to all Tfh cells), and the Tfh-cell subset by flow cytometry (BD FACSCanto™ II; BD Biosciences, USA). ..

Cytometry:

Article Title: Effect of CHK1 Inhibition on CPX-351 Cytotoxicity in vitro and ex vivo
Article Snippet: .. Analysis of cell cycle distribution and apoptosis After incubation for 24 h with CPX-351 in the absence or presence of CHK1 inhibitors, cells were resuspended in ice cold 0.1% (wt/vol) sodium citrate containing 50 µg/mL PI and 0.1% (wt/vol) Triton X-100, incubated at 4 °C overnight, and analyzed by flow microfluorimetry in the FL2 channel on a Becton Dickinson (Franklin Lakes, NJ) FASCanto II flow cytometer. .. After collection of 20,000 events, files were analyzed using Modfit (Verity Software, Topsham, ME) for cell cycle distribution or Becton Dickinson CellQuest software for events with < 2n DNA content.

Article Title: Aberrant populations of circulating T follicular helper cells and regulatory B cells underlying idiopathic pulmonary fibrosis
Article Snippet: .. We isolated heparinized peripheral blood mononuclear cells from fresh blood specimens by centrifugation over a discontinuous density gradient (Lympholyte-H; Cedarlane Laboratories Ltd., Canada) and investigated the proportion of Breg cells (to CD3− CD19+ B cells), Tfh cells (to CD3+ CD4+ T cells), PD-1+ ICOS+ Tfh cells (to all Tfh cells), and the Tfh-cell subset by flow cytometry (BD FACSCanto™ II; BD Biosciences, USA). ..

Article Title: NF-κB, p38 MAPK, ERK1/2, mTOR, STAT3 and increased glycolysis regulate stability of paricalcitol/dexamethasone-generated tolerogenic dendritic cells in the inflammatory environment
Article Snippet: .. Reagents and Abs Flow cytometry: commercial antibodies anti-CD86-FITC (clone 2231 FUN-1), CD274 (PD-L1)-FITC (clone MIH1), CD273 (PD-L2)-PE (clone MIH-18), HLA-DR-PE-Cy7 (clone L243), IFN-γ-FITC (clone 4SB3) were purchased from BD Biosciences; CD83-PerCP-Cy5.5 (clone HB15a) was purchased from Beckman Coulter; CD80-FITC (clone MAB104), CD40-PerCP-eFluor710 (clone 5C3), CD1a-PE-Cy7 (clone HI149) and CD4-PE-Cy7 (clone RPA-T4) were purchased from eBioscience; TLR2-FITC (clone T2.5), TIM-3-PE (clone F38-2E2), IL-10-PE (clone JES3-9D7), KI-67-PE (clone Ki-67) were purchased from BioLegend; CD14-PE-DL594 (clone MEM-15), CD11c-APC (clone BU15), CD3-AF700 (clone MEM-57), CD8-PE-Dy590 (clone MEM-31) were purchased from Exbio; CD85k (ILT-3)-PE (clone 293623), CD85d (IL-T4)-FITC (clone 287219) were purchased from R & D Systems. .. For western blot, anti-p-p38 MAPK, anti-p-ERK1/2, anti-p-JNK/SAPK, anti-p-IκB-α, anti-IDO, anti-p-mTOR, anti-p-STAT3, anti-p-p70S6K, anti-p38 MAPK, anti-ERK1/2, anti-JNK/SAPK and anti-STAT3 Ab were purchased from Cell Signaling Technology; anti-actin was from BioLegend.

Article Title: Absence of phagocyte NADPH oxidase 2 leads to severe inflammatory response in lungs of mice infected with Coccidioides
Article Snippet: .. In brief, absolute numbers of pulmonary leukocytes in the lung tissue preparations were enumerated by flow cytofluorometry using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ). .. Data were acquired with CellQuest Pro software (BD Biosciences) and analyzed using a FlowJo software package (Tree Star, Inc., Ashland, OR).

Incubation:

Article Title: Effect of CHK1 Inhibition on CPX-351 Cytotoxicity in vitro and ex vivo
Article Snippet: .. Analysis of cell cycle distribution and apoptosis After incubation for 24 h with CPX-351 in the absence or presence of CHK1 inhibitors, cells were resuspended in ice cold 0.1% (wt/vol) sodium citrate containing 50 µg/mL PI and 0.1% (wt/vol) Triton X-100, incubated at 4 °C overnight, and analyzed by flow microfluorimetry in the FL2 channel on a Becton Dickinson (Franklin Lakes, NJ) FASCanto II flow cytometer. .. After collection of 20,000 events, files were analyzed using Modfit (Verity Software, Topsham, ME) for cell cycle distribution or Becton Dickinson CellQuest software for events with < 2n DNA content.

Activation Assay:

Article Title: Selection of antitumor displayed peptides for the specific delivery of the anticancer drug lactaptin
Article Snippet: .. The exposure of phosphatidylserine (PS) at the outer surface of the cell membrane, and the activation of caspase-3 and −7 were analyzed by flow cytofluorometry with fluorescein isothiocyanate Annexin V Apoptosis Detection kit I (BD Pharmingen, San Jose, CA, USA) and the Vybrant FAM Caspase-3 and −7 assay kit (Life Technologies; Thermo Fisher Scientific, Inc.) using a FACSCanto II cell analyzer (BD Biosciences, Franklin Lakes, NJ, USA). .. Labeling of the recombinant lactaptin analogs with sulfo-Cy5 The recombinant lactaptin analogs RL2, Т1_RL and Т2_RL were isolated from E. coli and purified according to a protocol for the isolation of RL2 ( ).

Generated:

Article Title: Estrogenic Activity of Coumestrol, DDT, and TCDD in Human Cervical Cancer Cells
Article Snippet: .. Flow microfluorometry was performed and DNA histograms were generated and analyzed using a Becton Dickinson Flowscan (Franklin Lakes, NJ). .. This method correlates closely with other measures of apoptosis including TUNEL and Annexin V staining while providing additional cell cycle information [ ].

Expressing:

Article Title: IgE-FcεRI Interactions Determine HIV Coreceptor Usage and Susceptibility to Infection during Ontogeny of Mast Cells 1
Article Snippet: .. Analysis of the surface expression of CXCR4 and Fc ε RI-bound IgE on paraformaldehyde-treated prMC was performed by immunophenotyping and flow microfluorometry with a BD Biosciences FACSCalibur using CellQuest software as reported previously ( ). ..

FACS:

Article Title: Identification of a Boundary Domain Adjacent to the Potent Human Cytomegalovirus Enhancer That Represses Transcription of the Divergent UL127 Promoter †
Article Snippet: .. Fluorescence was measured by flow cytofluorometry (fluorescence-activated cell sorting [FACS]) using a FACScan (Becton Dickinson, Mountain View, Calif.). .. Uncompensated fluorescence values from FL1 (GFP fluorescence, 530-nm band pass filter) and FL2 (autofluorescence; a 585-nm band pass filter) were collected along with forward angle light scatter (FSC) and side scatter (SSC) for every event.

Centrifugation:

Article Title: Aberrant populations of circulating T follicular helper cells and regulatory B cells underlying idiopathic pulmonary fibrosis
Article Snippet: .. We isolated heparinized peripheral blood mononuclear cells from fresh blood specimens by centrifugation over a discontinuous density gradient (Lympholyte-H; Cedarlane Laboratories Ltd., Canada) and investigated the proportion of Breg cells (to CD3− CD19+ B cells), Tfh cells (to CD3+ CD4+ T cells), PD-1+ ICOS+ Tfh cells (to all Tfh cells), and the Tfh-cell subset by flow cytometry (BD FACSCanto™ II; BD Biosciences, USA). ..

Recombinase Polymerase Amplification:

Article Title: NF-κB, p38 MAPK, ERK1/2, mTOR, STAT3 and increased glycolysis regulate stability of paricalcitol/dexamethasone-generated tolerogenic dendritic cells in the inflammatory environment
Article Snippet: .. Reagents and Abs Flow cytometry: commercial antibodies anti-CD86-FITC (clone 2231 FUN-1), CD274 (PD-L1)-FITC (clone MIH1), CD273 (PD-L2)-PE (clone MIH-18), HLA-DR-PE-Cy7 (clone L243), IFN-γ-FITC (clone 4SB3) were purchased from BD Biosciences; CD83-PerCP-Cy5.5 (clone HB15a) was purchased from Beckman Coulter; CD80-FITC (clone MAB104), CD40-PerCP-eFluor710 (clone 5C3), CD1a-PE-Cy7 (clone HI149) and CD4-PE-Cy7 (clone RPA-T4) were purchased from eBioscience; TLR2-FITC (clone T2.5), TIM-3-PE (clone F38-2E2), IL-10-PE (clone JES3-9D7), KI-67-PE (clone Ki-67) were purchased from BioLegend; CD14-PE-DL594 (clone MEM-15), CD11c-APC (clone BU15), CD3-AF700 (clone MEM-57), CD8-PE-Dy590 (clone MEM-31) were purchased from Exbio; CD85k (ILT-3)-PE (clone 293623), CD85d (IL-T4)-FITC (clone 287219) were purchased from R & D Systems. .. For western blot, anti-p-p38 MAPK, anti-p-ERK1/2, anti-p-JNK/SAPK, anti-p-IκB-α, anti-IDO, anti-p-mTOR, anti-p-STAT3, anti-p-p70S6K, anti-p38 MAPK, anti-ERK1/2, anti-JNK/SAPK and anti-STAT3 Ab were purchased from Cell Signaling Technology; anti-actin was from BioLegend.

Software:

Article Title: IgE-FcεRI Interactions Determine HIV Coreceptor Usage and Susceptibility to Infection during Ontogeny of Mast Cells 1
Article Snippet: .. Analysis of the surface expression of CXCR4 and Fc ε RI-bound IgE on paraformaldehyde-treated prMC was performed by immunophenotyping and flow microfluorometry with a BD Biosciences FACSCalibur using CellQuest software as reported previously ( ). ..

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  • 92
    Becton Dickinson flow cytometry measurements
    The viability of the L. amazonensis promastigotes was analyzed by labeling with propidium iodide (PI). Parasites cultured at 26, 34, or 37°C for 1, 2, 4, or 24 h were stained with PI and analyzed by flow <t>cytometry.</t> NC, negative control (fresh parasites); PC, positive control (parasites treated with 20 mM H 2 O 2 for 1 h). The bars denote the average of three measurements, and the error bars denote the SD. ANOVA was performed, followed by a post hoc Tukey’s test ( ∗∗∗ P
    Flow Cytometry Measurements, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flow cytometry measurements/product/Becton Dickinson
    Average 92 stars, based on 179 article reviews
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    99
    Becton Dickinson flow cytometry analysis
    Expression of SAF-1 and SAF-3 variants driven by alternative promoters The biochromatic fluorescent reporters were driven by −1692/+277 promoter of SAF/MAZ gene or −1401/−277, −595/+277, and −1692/+277Δ−1401/−595 SAF-1. The average FIs of cells were analyzed for SAF-1 and SAF-3 by flow <t>cytometry</t> ( A ).The data shown represent the difference of mean ±SEM of three separate expreiments between two groups indicated by line below star symbols (** P
    Flow Cytometry Analysis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 4595 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flow cytometry analysis/product/Becton Dickinson
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    Becton Dickinson facscalibur flow cytometer
    Vanillic acid inhibits the proliferation of HCT116 cells via blocking cell cycle progression in the G1 phase. ( A ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Subsequently, cell cycle status was analyzed by by <t>FACSCalibur</t> flow cytometry. ( B ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Cyclin D1 and c-Myc were detected by Western blot analysis. Levels of tubulin were used as a loading control. ( C ) Data were shown as mean ± SD ( n = 3). ** p
    Facscalibur Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 14797 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/facscalibur flow cytometer/product/Becton Dickinson
    Average 99 stars, based on 14797 article reviews
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    93
    Becton Dickinson flow cytometric detection
    Splenic B1 cells correlate with T cell exhaustion during chronic SIV-infection. (A) Flow <t>cytometric</t> analysis of chronically SIV-infected splenic exhausted T cells by gating on CD4 + CD6 + PD-1 + and CD8 + CD6 + PD-1 + respectively. Data are representative of 25 macaques. For CD4 + T cells: CD6 + PD-1 − (non-exhausted), CD6 + PD-1 lo (exhausted), and CD6 + PD-1 hi (terminally exhausted). For CD8 + T cells: CD6 + PD-1 − (non-exhausted) and CD6 + PD-1 + (exhausted). (B) The correlation of splenic B1 cells with terminally exhausted CD4 + T cells. (C) The correlation of splenic B1 cells with exhausted splenic CD8 + T cells. (D) The correlation of splenic CD11b + B1 cells with terminally exhausted CD4 + T cells. Data in (B–D) are from 25 macaques. For statistical analysis, non-parametric Spearman rank correlations were performed. All tests were two-tailed.
    Flow Cytometric Detection, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The viability of the L. amazonensis promastigotes was analyzed by labeling with propidium iodide (PI). Parasites cultured at 26, 34, or 37°C for 1, 2, 4, or 24 h were stained with PI and analyzed by flow cytometry. NC, negative control (fresh parasites); PC, positive control (parasites treated with 20 mM H 2 O 2 for 1 h). The bars denote the average of three measurements, and the error bars denote the SD. ANOVA was performed, followed by a post hoc Tukey’s test ( ∗∗∗ P

    Journal: Frontiers in Microbiology

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells

    doi: 10.3389/fmicb.2018.03056

    Figure Lengend Snippet: The viability of the L. amazonensis promastigotes was analyzed by labeling with propidium iodide (PI). Parasites cultured at 26, 34, or 37°C for 1, 2, 4, or 24 h were stained with PI and analyzed by flow cytometry. NC, negative control (fresh parasites); PC, positive control (parasites treated with 20 mM H 2 O 2 for 1 h). The bars denote the average of three measurements, and the error bars denote the SD. ANOVA was performed, followed by a post hoc Tukey’s test ( ∗∗∗ P

    Article Snippet: Flow cytometry measurements were performed on a BD FACS Accuri C6 flow cytometer (BD Biosciences) and the data evaluated by FCAP ArrayTM software (BD Bioscience).

    Techniques: Labeling, Cell Culture, Staining, Flow Cytometry, Cytometry, Negative Control, Positive Control

    Expression of SAF-1 and SAF-3 variants driven by alternative promoters The biochromatic fluorescent reporters were driven by −1692/+277 promoter of SAF/MAZ gene or −1401/−277, −595/+277, and −1692/+277Δ−1401/−595 SAF-1. The average FIs of cells were analyzed for SAF-1 and SAF-3 by flow cytometry ( A ).The data shown represent the difference of mean ±SEM of three separate expreiments between two groups indicated by line below star symbols (** P

    Journal: Bioscience Reports

    Article Title: Revealing the alternative promoter usage of SAF/MAZ gene by bichromatic fluorescent reporter construct

    doi: 10.1042/BSR20171668

    Figure Lengend Snippet: Expression of SAF-1 and SAF-3 variants driven by alternative promoters The biochromatic fluorescent reporters were driven by −1692/+277 promoter of SAF/MAZ gene or −1401/−277, −595/+277, and −1692/+277Δ−1401/−595 SAF-1. The average FIs of cells were analyzed for SAF-1 and SAF-3 by flow cytometry ( A ).The data shown represent the difference of mean ±SEM of three separate expreiments between two groups indicated by line below star symbols (** P

    Article Snippet: Although there were statistically meaningful differences amongst these four constructs for SAF-1 expression by flow cytometry analysis, the change of SAF-1 expression was not as obvious as for SAF-3 expression ( A) .

    Techniques: Expressing, Flow Cytometry, Cytometry

    Repression of SAF-1 and SAF-3 promoter by transcription factor Elk-1 and endogenous SAF-1/SAF-3 expression Elk-1 cis -element on SAF/MAZ promoter was identified by EMSA ( A ). Horizontal black and red arrows represent the specific protein/DNA binding bands and anti-His/His tagged ELK-1/DNA probe supershift bands, respectively. The bichromatic fluorescent reporter plasmids were transiently co-transfected with empty plasmid, pCGN-Elk-1 and pN3-Sp1 into HeLa cells. The SAF-1 and SAF-3 promoter activation status were tested by either laser co-focus microscopy ( B ) or by flow cytometry analysis ( C ). The endogenous SAF-1 and SAF-3 mRNA expression status in HeLa cells are shown by ( D ). In (D), lanes 1 and 8 are DNA markers, lanes 2 and 5 are SAF-1 mRNA levels (271 bp), lanes 3 and 6 are SAF-3 mRNA levels (208 bp), and lanes 4 and 7 are GAPDH mRNA levels (177 bp). Abbreviation: EMSA, electrophoretic mobility shift assay.

    Journal: Bioscience Reports

    Article Title: Revealing the alternative promoter usage of SAF/MAZ gene by bichromatic fluorescent reporter construct

    doi: 10.1042/BSR20171668

    Figure Lengend Snippet: Repression of SAF-1 and SAF-3 promoter by transcription factor Elk-1 and endogenous SAF-1/SAF-3 expression Elk-1 cis -element on SAF/MAZ promoter was identified by EMSA ( A ). Horizontal black and red arrows represent the specific protein/DNA binding bands and anti-His/His tagged ELK-1/DNA probe supershift bands, respectively. The bichromatic fluorescent reporter plasmids were transiently co-transfected with empty plasmid, pCGN-Elk-1 and pN3-Sp1 into HeLa cells. The SAF-1 and SAF-3 promoter activation status were tested by either laser co-focus microscopy ( B ) or by flow cytometry analysis ( C ). The endogenous SAF-1 and SAF-3 mRNA expression status in HeLa cells are shown by ( D ). In (D), lanes 1 and 8 are DNA markers, lanes 2 and 5 are SAF-1 mRNA levels (271 bp), lanes 3 and 6 are SAF-3 mRNA levels (208 bp), and lanes 4 and 7 are GAPDH mRNA levels (177 bp). Abbreviation: EMSA, electrophoretic mobility shift assay.

    Article Snippet: Although there were statistically meaningful differences amongst these four constructs for SAF-1 expression by flow cytometry analysis, the change of SAF-1 expression was not as obvious as for SAF-3 expression ( A) .

    Techniques: Expressing, Binding Assay, Transfection, Plasmid Preparation, Activation Assay, Microscopy, Flow Cytometry, Cytometry, Electrophoretic Mobility Shift Assay

    Vanillic acid inhibits the proliferation of HCT116 cells via blocking cell cycle progression in the G1 phase. ( A ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Subsequently, cell cycle status was analyzed by by FACSCalibur flow cytometry. ( B ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Cyclin D1 and c-Myc were detected by Western blot analysis. Levels of tubulin were used as a loading control. ( C ) Data were shown as mean ± SD ( n = 3). ** p

    Journal: International Journal of Molecular Sciences

    Article Title: Vanillic Acid Suppresses HIF-1α Expression via Inhibition of mTOR/p70S6K/4E-BP1 and Raf/MEK/ERK Pathways in Human Colon Cancer HCT116 Cells

    doi: 10.3390/ijms20030465

    Figure Lengend Snippet: Vanillic acid inhibits the proliferation of HCT116 cells via blocking cell cycle progression in the G1 phase. ( A ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Subsequently, cell cycle status was analyzed by by FACSCalibur flow cytometry. ( B ) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Cyclin D1 and c-Myc were detected by Western blot analysis. Levels of tubulin were used as a loading control. ( C ) Data were shown as mean ± SD ( n = 3). ** p

    Article Snippet: The DNA content was analyzed using a FACSCalibur flow cytometer with Cell Quest software (Becton-Dickinson, Franklin Lakes, NJ, USA).

    Techniques: Blocking Assay, Concentration Assay, Cell Culture, Flow Cytometry, Cytometry, Western Blot

    Splenic B1 cells correlate with T cell exhaustion during chronic SIV-infection. (A) Flow cytometric analysis of chronically SIV-infected splenic exhausted T cells by gating on CD4 + CD6 + PD-1 + and CD8 + CD6 + PD-1 + respectively. Data are representative of 25 macaques. For CD4 + T cells: CD6 + PD-1 − (non-exhausted), CD6 + PD-1 lo (exhausted), and CD6 + PD-1 hi (terminally exhausted). For CD8 + T cells: CD6 + PD-1 − (non-exhausted) and CD6 + PD-1 + (exhausted). (B) The correlation of splenic B1 cells with terminally exhausted CD4 + T cells. (C) The correlation of splenic B1 cells with exhausted splenic CD8 + T cells. (D) The correlation of splenic CD11b + B1 cells with terminally exhausted CD4 + T cells. Data in (B–D) are from 25 macaques. For statistical analysis, non-parametric Spearman rank correlations were performed. All tests were two-tailed.

    Journal: Frontiers in Immunology

    Article Title: A Pathogenic Role for Splenic B1 Cells in SIV Disease Progression in Rhesus Macaques

    doi: 10.3389/fimmu.2019.00511

    Figure Lengend Snippet: Splenic B1 cells correlate with T cell exhaustion during chronic SIV-infection. (A) Flow cytometric analysis of chronically SIV-infected splenic exhausted T cells by gating on CD4 + CD6 + PD-1 + and CD8 + CD6 + PD-1 + respectively. Data are representative of 25 macaques. For CD4 + T cells: CD6 + PD-1 − (non-exhausted), CD6 + PD-1 lo (exhausted), and CD6 + PD-1 hi (terminally exhausted). For CD8 + T cells: CD6 + PD-1 − (non-exhausted) and CD6 + PD-1 + (exhausted). (B) The correlation of splenic B1 cells with terminally exhausted CD4 + T cells. (C) The correlation of splenic B1 cells with exhausted splenic CD8 + T cells. (D) The correlation of splenic CD11b + B1 cells with terminally exhausted CD4 + T cells. Data in (B–D) are from 25 macaques. For statistical analysis, non-parametric Spearman rank correlations were performed. All tests were two-tailed.

    Article Snippet: Flow Cytometric Detection of IL-10 IL-10 staining was performed by culturing splenocytes from chronically SIV-infected macaques in complete media in the presence of BD Golgistop (1 μl; BD) containing monensin for 4 h prior to cell surface staining.

    Techniques: Infection, Flow Cytometry, Two Tailed Test

    B1 cells possess T-cell exhaustion-inducing potential. (A) Representative example of flow cytometric analysis of PD-L1 and PD-L2 expression on splenic B1 cells by gating on CD3 − CD19 + CD43 + CD27 + CD11b + or CD3 − CD19 + CD43 + CD27 + CD11b − cells. Shown to the right are mean frequencies ± SEM. Data are from 8 macaques. (B) Representative example of flow cytometric analysis of IL-10 + splenic B1 cells by gating on CD3 − CD19 + CD43 + CD27 + CD11b + or CD3 − CD19 + CD43 + CD27 + CD11b − cells. Shown to the right are mean frequencies ± SEM. Data are from 4 macaques. Splenocytes were acquired during the chronic phase of SIV infection. For statistical analysis nonparametric unpaired Mann–Whitney tests were performed. *** p

    Journal: Frontiers in Immunology

    Article Title: A Pathogenic Role for Splenic B1 Cells in SIV Disease Progression in Rhesus Macaques

    doi: 10.3389/fimmu.2019.00511

    Figure Lengend Snippet: B1 cells possess T-cell exhaustion-inducing potential. (A) Representative example of flow cytometric analysis of PD-L1 and PD-L2 expression on splenic B1 cells by gating on CD3 − CD19 + CD43 + CD27 + CD11b + or CD3 − CD19 + CD43 + CD27 + CD11b − cells. Shown to the right are mean frequencies ± SEM. Data are from 8 macaques. (B) Representative example of flow cytometric analysis of IL-10 + splenic B1 cells by gating on CD3 − CD19 + CD43 + CD27 + CD11b + or CD3 − CD19 + CD43 + CD27 + CD11b − cells. Shown to the right are mean frequencies ± SEM. Data are from 4 macaques. Splenocytes were acquired during the chronic phase of SIV infection. For statistical analysis nonparametric unpaired Mann–Whitney tests were performed. *** p

    Article Snippet: Flow Cytometric Detection of IL-10 IL-10 staining was performed by culturing splenocytes from chronically SIV-infected macaques in complete media in the presence of BD Golgistop (1 μl; BD) containing monensin for 4 h prior to cell surface staining.

    Techniques: Flow Cytometry, Expressing, Infection, MANN-WHITNEY