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Becton Dickinson facsaria flow cytometer
Facsaria 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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facsaria flow cytometer - by Bioz Stars, 2021-05
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Flow Cytometry:

Article Title: Synergistic STING activation by PC7A nanovaccine and ionizing radiation improves cancer immunotherapy.
Article Snippet: Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity. .. Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity. .. Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity.

Article Title: NK Cells Inhibit T-bet-deficient, Autoreactive Th17 Cells
Article Snippet: After 18–24 h, BrdU-labelled cells harvested from the culture or single cell suspensions prepared from spleen were stained with mAb against CD4, IL-17 and BrdU. .. Samples were analysed on a FACSAria™ flow cytometer using Diva™ software. .. Western blots were performed using anti-pSTAT3 (49/p-Stat3) and anti-STAT3 (84/Stat3) (BD Bioscience) antibodies followed by HRP-conjugated goat anti-mouse IgG and ECL detection system (Pierce, Rockford, IL, USA).

Article Title: Synergistic effect of the herbal mixture C5E on gemcitabine treatment in PANC-1 cells
Article Snippet: Following the incubation, the cells were centrifuged (250 × g; 5 min; 25°C) and resuspended in ice-cold HBSS (Welgene, Inc.) containing 2% FBS and 10 mM HEPES buffer. .. The cells were then analyzed using a FACSAria™ flow cytometer (BD Biosciences). .. Hoechst 33342 was excited at 357 nm, and its fluorescence was analyzed using a dual-wavelength (blue, 402–446 nm; red, 650–670 nm).

Article Title: Expression of the progenitor marker NG2/CSPG4 predicts poor survival and resistance to ionising radiation in glioblastoma
Article Snippet: After two washes in PermWash solution, cells were suspended in PBS supplemented with 0.5% BSA and stained with Hoechst solution at 0.1 μg/ml in order to discriminate nuclear cells from debris. .. The samples were run on a FACSAria™ flow cytometer and analysed with FACSDiva™ software (BD Biosciences). .. Diagrams were then created using FlowJo flow software version 7.2.5 (Tree Star Inc, Ashland, OR, USA).

Article Title: Expansion of regulatory T cells via IL-2/anti-IL-2 mAb complexes suppresses experimental myasthenia
Article Snippet: For intracellular Foxp3 expression, 106 cells were stained for surface molecules with anti-CD4-FITC (H129.19) and anti-CD25-allophycocyanin (PC61.5), fixed and permeabilized, and then stained for intracellular molecules with anti-Foxp3-PE (FJK-16s) (eBioscience). .. Samples were read using a FACSAria™ flow cytometer and data analyzed using Diva™ software. .. To label the cultured cells, 10 µL of 1mM BrdU was added to 1mL of culture media.

Article Title: A novel histone H4 variant H4G regulates rDNA transcription in breast cancer
Article Snippet: Cell-cycle analysis MCF7 cells were synchronized by serum starvation for 24 h and induced to re-enter the cell cycle by the addition of serum. .. Cells were then harvested for propidium iodide staining and analyzed by fluorescence-activated cell sorting (FACS) using a Becton Dickinson FACSAria™ III flow cytometer (BD Biosciences) to determine the cell-cycle fraction. .. Data were analyzed using FCS Express software (De Novo Software).

Article Title: Molecular Basis for Autosomal-Dominant Renal Fanconi Syndrome Caused by HNF4A
Article Snippet: Lentiviruses were diluted 1:100 to 1:1000 in MEFM containing 8μg mL-1 polybrene (Santa Cruz) and transduced for 12 h on 6 consecutive days. .. Reprogrammed, GFP-positive cells were sorted 14 days after the last viral transduction using a BD FACSAria™ Fusion flow cytometer (Becton Dickinson). .. For immunocytochemistry, cells were washed three times in PBS, fixed for 20 min in 4% paraformaldehyde in PBS, blocked for 10 min in PBS + 3% BSA + 0,1% Tween + 0,1% Triton and incubated overnight at 4°C with primary antibodies diluted in PBS + 3% BSA + 0,1% tween + 0,1% triton.

Article Title: An Oral FMT Capsule as Efficient as an Enema for Microbiota Reconstruction Following Disruption by Antibiotics, as Assessed in an In Vitro Human Gut Model
Article Snippet: Bacterial suspensions were thus incubated for 15 min at room temperature in the dark with 3.3 mM SYTO 9 and 0.375 mM Propidium Iodide and transferred into BD Trucount™ Tubes (BD Biosciences, San Jose, CA, USA). .. Flow cytometry analysis was performed on a BD™ LSR II cytometer (BD Biosciences, San Jose, CA, USA) and data were collected with BD FACSDivaTM software. ..

Software:

Article Title: Synergistic STING activation by PC7A nanovaccine and ionizing radiation improves cancer immunotherapy.
Article Snippet: Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity. .. Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity. .. Solid cancers are able to escape immune surveillance and are resistant to current treatment in immunotherapy.. Recent evidence indicates the critical role of the stimulator of interferon genes (STING) pathway in antitumor immunity.

Article Title: NK Cells Inhibit T-bet-deficient, Autoreactive Th17 Cells
Article Snippet: After 18–24 h, BrdU-labelled cells harvested from the culture or single cell suspensions prepared from spleen were stained with mAb against CD4, IL-17 and BrdU. .. Samples were analysed on a FACSAria™ flow cytometer using Diva™ software. .. Western blots were performed using anti-pSTAT3 (49/p-Stat3) and anti-STAT3 (84/Stat3) (BD Bioscience) antibodies followed by HRP-conjugated goat anti-mouse IgG and ECL detection system (Pierce, Rockford, IL, USA).

Article Title: Expression of the progenitor marker NG2/CSPG4 predicts poor survival and resistance to ionising radiation in glioblastoma
Article Snippet: After two washes in PermWash solution, cells were suspended in PBS supplemented with 0.5% BSA and stained with Hoechst solution at 0.1 μg/ml in order to discriminate nuclear cells from debris. .. The samples were run on a FACSAria™ flow cytometer and analysed with FACSDiva™ software (BD Biosciences). .. Diagrams were then created using FlowJo flow software version 7.2.5 (Tree Star Inc, Ashland, OR, USA).

Article Title: Expansion of regulatory T cells via IL-2/anti-IL-2 mAb complexes suppresses experimental myasthenia
Article Snippet: For intracellular Foxp3 expression, 106 cells were stained for surface molecules with anti-CD4-FITC (H129.19) and anti-CD25-allophycocyanin (PC61.5), fixed and permeabilized, and then stained for intracellular molecules with anti-Foxp3-PE (FJK-16s) (eBioscience). .. Samples were read using a FACSAria™ flow cytometer and data analyzed using Diva™ software. .. To label the cultured cells, 10 µL of 1mM BrdU was added to 1mL of culture media.

Article Title: An Oral FMT Capsule as Efficient as an Enema for Microbiota Reconstruction Following Disruption by Antibiotics, as Assessed in an In Vitro Human Gut Model
Article Snippet: Bacterial suspensions were thus incubated for 15 min at room temperature in the dark with 3.3 mM SYTO 9 and 0.375 mM Propidium Iodide and transferred into BD Trucount™ Tubes (BD Biosciences, San Jose, CA, USA). .. Flow cytometry analysis was performed on a BD™ LSR II cytometer (BD Biosciences, San Jose, CA, USA) and data were collected with BD FACSDivaTM software. ..

Cytometry:

Article Title: NK Cells Inhibit T-bet-deficient, Autoreactive Th17 Cells
Article Snippet: After 18–24 h, BrdU-labelled cells harvested from the culture or single cell suspensions prepared from spleen were stained with mAb against CD4, IL-17 and BrdU. .. Samples were analysed on a FACSAria™ flow cytometer using Diva™ software. .. Western blots were performed using anti-pSTAT3 (49/p-Stat3) and anti-STAT3 (84/Stat3) (BD Bioscience) antibodies followed by HRP-conjugated goat anti-mouse IgG and ECL detection system (Pierce, Rockford, IL, USA).

Article Title: Expansion of regulatory T cells via IL-2/anti-IL-2 mAb complexes suppresses experimental myasthenia
Article Snippet: For intracellular Foxp3 expression, 106 cells were stained for surface molecules with anti-CD4-FITC (H129.19) and anti-CD25-allophycocyanin (PC61.5), fixed and permeabilized, and then stained for intracellular molecules with anti-Foxp3-PE (FJK-16s) (eBioscience). .. Samples were read using a FACSAria™ flow cytometer and data analyzed using Diva™ software. .. To label the cultured cells, 10 µL of 1mM BrdU was added to 1mL of culture media.

Article Title: An Oral FMT Capsule as Efficient as an Enema for Microbiota Reconstruction Following Disruption by Antibiotics, as Assessed in an In Vitro Human Gut Model
Article Snippet: Bacterial suspensions were thus incubated for 15 min at room temperature in the dark with 3.3 mM SYTO 9 and 0.375 mM Propidium Iodide and transferred into BD Trucount™ Tubes (BD Biosciences, San Jose, CA, USA). .. Flow cytometry analysis was performed on a BD™ LSR II cytometer (BD Biosciences, San Jose, CA, USA) and data were collected with BD FACSDivaTM software. ..

Staining:

Article Title: A novel histone H4 variant H4G regulates rDNA transcription in breast cancer
Article Snippet: Cell-cycle analysis MCF7 cells were synchronized by serum starvation for 24 h and induced to re-enter the cell cycle by the addition of serum. .. Cells were then harvested for propidium iodide staining and analyzed by fluorescence-activated cell sorting (FACS) using a Becton Dickinson FACSAria™ III flow cytometer (BD Biosciences) to determine the cell-cycle fraction. .. Data were analyzed using FCS Express software (De Novo Software).

Fluorescence:

Article Title: A novel histone H4 variant H4G regulates rDNA transcription in breast cancer
Article Snippet: Cell-cycle analysis MCF7 cells were synchronized by serum starvation for 24 h and induced to re-enter the cell cycle by the addition of serum. .. Cells were then harvested for propidium iodide staining and analyzed by fluorescence-activated cell sorting (FACS) using a Becton Dickinson FACSAria™ III flow cytometer (BD Biosciences) to determine the cell-cycle fraction. .. Data were analyzed using FCS Express software (De Novo Software).

FACS:

Article Title: A novel histone H4 variant H4G regulates rDNA transcription in breast cancer
Article Snippet: Cell-cycle analysis MCF7 cells were synchronized by serum starvation for 24 h and induced to re-enter the cell cycle by the addition of serum. .. Cells were then harvested for propidium iodide staining and analyzed by fluorescence-activated cell sorting (FACS) using a Becton Dickinson FACSAria™ III flow cytometer (BD Biosciences) to determine the cell-cycle fraction. .. Data were analyzed using FCS Express software (De Novo Software).

Transduction:

Article Title: Molecular Basis for Autosomal-Dominant Renal Fanconi Syndrome Caused by HNF4A
Article Snippet: Lentiviruses were diluted 1:100 to 1:1000 in MEFM containing 8μg mL-1 polybrene (Santa Cruz) and transduced for 12 h on 6 consecutive days. .. Reprogrammed, GFP-positive cells were sorted 14 days after the last viral transduction using a BD FACSAria™ Fusion flow cytometer (Becton Dickinson). .. For immunocytochemistry, cells were washed three times in PBS, fixed for 20 min in 4% paraformaldehyde in PBS, blocked for 10 min in PBS + 3% BSA + 0,1% Tween + 0,1% Triton and incubated overnight at 4°C with primary antibodies diluted in PBS + 3% BSA + 0,1% tween + 0,1% triton.

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    Becton Dickinson cd74
    <t>CD74</t> −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p
    Cd74, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd74/product/Becton Dickinson
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd74 - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    86
    Becton Dickinson facsaria flow cytometer
    Activation markers on CD4+ and CD8+ T cell subsets upon T. cruzi and ribosomal protein activation. PBMC isolated from patients with chronic Chagas' disease Cardiomyopathy (CCC; n = 27) and non-infected individuals (NI; n = 20) were seeded at 2.5×10 6 cells/well and stimulated with T. cruzi lysate, P2β or CP0 proteins (10 µg/ml) or medium alone for 6 days. PBMC were stained with CD3-APC, CD4-PE-Cy5 or CD8-PE-Cy5 and activation marker-specific labeled antibodies (CD25-FITC and HLA-DR-PE) prior to flow cytometry analysis. 10,000–15,000 events in the lymphocyte gate (R1 gate) were acquired using a <t>FACSAria</t> flow cytometer (Becton Dickinson); dead cells were excluded by forward vs side-scatter (FSC/SSC) gating. A) Gate-pathway used to determine the activation expression in the populations graphed in B. B) Results were expressed as the percentage of CD25+ or HLA-DR+ cells in CD3+CD4+ (R2 gate) or CD3+CD8+ (R3 gate) lymphocytes. Horizontal lines represent the median and percentiles 25–75th, vertical lines represent percentiles 5–95th. Statistical analysis was performed using the Mann-Whitney U Test, *** P
    Facsaria 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
    https://www.bioz.com/result/facsaria flow cytometer/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    facsaria flow cytometer - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

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    CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 −/− HSPCs show a higher potential to repopulate the BM. Lethally irradiated WT CD45.1 recipient mice were reconstituted with 7.5*10 4 sorted LSK cells from WT (CD45.1), and 7.5 × 10 4 sorted LSK from CD74 −/− (CD45.2) at a 1:1 ratio. Percent of donor-derived cells was analyzed in the BM after 6 and 18 weeks. (A) Total BM cells; Data A in S4 Data (B) myeloid cells; Data B in S4 Data (C) immature BM B cells; Data C in S4 Data and (D) mature BM B cells, Data D in S4 Data . (E) Percent of donor-derived cells was analyzed in LSK and CD34-LSK cells 18 weeks posttransplant. n = 6–8. Data E in S4 Data . n = 6–8. Bars show SEM. Unpaired two-tailed t test * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Irradiation, Mouse Assay, Derivative Assay, Two Tailed Test

    CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 −/− HSPCs demonstrate enhanced long-term self-renewal capacity. (A–F) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 3:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 3:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (A) Total BM cells; Data A in S5 Data (B) BM myeloid cells; Data B in S5 Data (C) BM immature B cells; Data C in S5 Data (D) LSK; Data D in S5 Data (E) CD34-/LSK; Data E in S5 Data (F) Mature BM B cells; Data F in S5 Data . n = 13. (G–L) Lethally irradiated WT(CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT(CD45.2) at a 9:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 9:1 ratio. Mice were analyzed 16 weeks after transplantation. Graphs show percent of donor-derived cells from both WTCD45.1/WTCD45.2 and WTCD45.1/CD74 −/− CD45.2 chimera. (G) Total BM cells; Data G in S5 Data (H) BM myeloid cells; Data H in S5 Data (I) BM immature B cells; Data I in S5 Data (J) LSK; Data J in S5 Data (K) CD34-/LSK; Data K in S5 Data (L) Mature BM B cells; Data L in S5 Data . n = 12. Bars show SEM. Unpaired two-tailed t test * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Irradiation, Mouse Assay, Derivative Assay, Transplantation Assay, Two Tailed Test

    Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: Expansion of HSPCs in the BM of CD74 −/− mice. (A) BM cells derived from WT or CD74 −/− were purified. Histograms show representative analysis of CD74 expression on HSPCs in WT and CD74 −/− mice. n = 3. (B) Total BM cellularity per femur and tibia in WT and CD74 −/− mice, Data A in S1 Data . (C–J) The percent of the different populations in WT and CD74 −/− -derived BM cells. (C) Lin-; Data B in S1 Data (D) Representative FACS analysis of WT and CD74 −/− HSPCs; (E) LSK; Data C in S1 Data (F) CD34-/LSK; Data D in S1 Data and (G) CD34+; Data E in S1 Data (H) CD150+CD48-LSK; Data F in S1 Data (I) CD150-CD48-/LSK; Data G in S1 Data and (J) CD150-CD48+/LSK; n = 14–18, Data H in S1 Data . (K) CFUC assay: Total BM cells from WT and CD74 −/− mice were seeded at 15,000 cells/mL in semisolid cultures supplemented with cytokines and nutrients. CFU-C were counted 7 days later; n = 7, Data I in S1 Data . Bars show SEM. Unpaired two-tailed t test * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Mouse Assay, Derivative Assay, Purification, Expressing, FACS, Two Tailed Test

    CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 −/− HSPCs have an advantage in BM repopulation. Lethally irradiated WT (CD45.1) mice were transplanted with BM derived from WT (CD45.1) and WT (CD45.2) at a 1:1 ratio, or BM derived from WT (CD45.1) and CD74 −/− (CD45.2) mice at a 1:1 ratio. (A) Representative BM FACS staining. Percent of donor-derived cells was analyzed in the BM after 6, 16, and 24 weeks in (B) Total BM cells; Data A in S3 Data (C) myeloid cells (CD11B+); Data B in S3 Data (D) B cells (B220+); Data C in S3 Data (E) LSK; Data D in S3 Data (F) CD34-/LSK; Data E in S3 Data (G) immature BM B cells (B220+IgD-); Data F in S3 Data (H) mature BM B cells (B220+ IgM+ IgD+); Data G in S3 Data . n = 8–18. Bars show SEM. Unpaired two-tailed t test * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Irradiation, Mouse Assay, Derivative Assay, FACS, Staining, Two Tailed Test

    CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 can serve as a potential target for therapy. (A, B) WT and CD74 −/− BM cells were cultured alone or incubated with blocking anti-CD74 antibody (20, 50, and 100 μg/ml). After 48 h, percent LSK from live cells was analyzed by FACS; n = 4–7, Data A and B in S8 Data . (C, D) Survival curve: 5-FU (150 mg/kg and 125mg/kg) was injected to WT and CD74 −/− mice once a week. Log-rank test *

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Cell Culture, Incubation, Blocking Assay, FACS, Injection, Mouse Assay

    Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: Accumulation of HSPCs is not CXCR4 dependent. (A, B) FACS analysis of CXCR4 expression on BM LSK and BM CD34-/LSK of WT and CD74 −/− mice, n = 7, Data A in S6 Data . Representative histograms are shown. (C–E) FACS analysis for HSPCs in the PB of WT and CD74 −/− . (C) Dot plot analysis of LSK in WT and CD74 −/− mice. (D, E) Cell number of (D) LSK and (E) CD34-LSK in 600 μl blood. WT n = 6 CD74 −/− n = 7, Data B and C in S6 Data . (F) AMD3100 (20 mg/kg −1 ) was injected to WT and CD74 −/− mice. After 2 h, percent of LSK in the PB was analyzed; n = 9–11, Data D in S6 Data . (G–I) FACS staining of WT and CD74 −/− HSPCs for Ki-67. Results are presented as: (G) percent of CD34-/LSK Ki-67 and CD34-/LSK Ki-67+ from total BM cells, Data E in S6 Data ; (H) percent of CD34+/LSK Ki-67- and CD34+/LSK Ki-67+ from total BM cells, Data F in S6 Data ; and (I) percent of Ki-67+ from CD34-/LSK and percent of Ki-67+ from CD34+LSK, n = 15, Data G in S6 Data . (J, K) Mice were fed with 0.8 mg/ml BrdU in their drinking water for 3 days, and BrdU incorporation was analyzed by FACS. Results are represented as: (J) percent of LSK BrdU- and LSK BrdU+ from total BM cells, Data H in S6 Data ; (K) percent of BrdU+ in LSK; n = 12–14, Data I in S6 Data . Bars show SEM. Unpaired two-tailed t test: * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: FACS, Expressing, Mouse Assay, Injection, Staining, BrdU Incorporation Assay, Two Tailed Test

    CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 −/− HSPC expansion is cell intrinsic. Lethally irradiated WT or CD74 −/− mice were transplanted with either WT or CD74 −/− total BM cells. Long-term reconstitution was evaluated 16 weeks posttransplantation. Percent of total BM cells was calculated for (A) LIN -; Data A in S2 Data (B) LSK; Data B in S2 Data and (C) CD34-/LSK; Data C in S2 Data n = 5–12. Bars show SEM. Unpaired two-tailed t test *

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Irradiation, Mouse Assay, Two Tailed Test

    CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

    Journal: PLoS Biology

    Article Title: CD74 is a regulator of hematopoietic stem cell maintenance

    doi: 10.1371/journal.pbio.3001121

    Figure Lengend Snippet: CD74 regulates the survival of HSPCs and CD18 expression. (A, B) FACS staining of WT and CD74-deficient HSPCs for ROS. (A) Results are presented as the number of ROS high cells per 10 6 cells, n = 9, Data A in S7 Data . (B) Percentage of ROS high in LSK, Data B in S7 Data . (C, D) Percent of LSK (C) Data C in S7 Data , and CD34- (D) after 6 days of NAC injections (50 mg kg −1 ); n = 5, Data D in S7 Data . (E, F) FACS analysis of HSPCs from WT and CD74 −/− mice for Annexin V (E); n = 10–12, Data E in S7 Data , and after 24 h under hypoxic (F); n = 3 (each dot represents a duplicate determination), Data F in S7 Data . (G) Ratio of Annexin V+ CD74 −/− to WT of HSPCs under hypoxic and normoxic conditions, Data G in S7 Data . (H, I) FACS analysis of HSCs from WT and CD74 −/− mice for HIF-1α; n = 7–8, Data H in S7 Data . (J) Sorted WT and CD74 −/− CD34-/LSK cells were analyzed for CD18 mRNA levels; n = 3. The bars show the DESeq2 normalized counts for the CD18 gene, Data I in S7 Data . (K) Binding of CD74–ICD to CD18 promoter and intron regions in Lin− samples. ChIP-seq analysis using anti-CD74 antibody. (L) FACS analysis of HSCs from WT and CD74 −/− mice for CD18. Graph summarizes the results of 6 mice in each group, Data J in S7 Data . (M) FACS analysis of HSCs from WT and MIF −/− mice for CD18; n = 3, Data K in S7 Data . (N) WT and CD74 −/− BM were cultured with or without the MIF inhibitor, ISO-1, for 48 h, percent CD18 on CD34-/LSK was analyzed by FACS; n = 6, Data L in S7 Data . (O) WT (CD45.1) Lin negative cells were cultured in the presence of WT (CD45.2) total BM or MIF −/− (CD45.2) total BM for 48 h. The percent CD18 on CD34-/LSK cells (CD45.1) was analyzed by FACS; n = 8, Data M in S7 Data . Bars show SEM. Unpaired two-tailed t test * p

    Article Snippet: Generation of chimeric mice For the microenvironment experiment: Lethally irradiated (950 Rad) C57BL/6 (WT) recipient mice were reconstituted with 5*106 WT or CD74−/− BM cells.

    Techniques: Expressing, FACS, Staining, Mouse Assay, Binding Assay, Chromatin Immunoprecipitation, Cell Culture, Two Tailed Test

    Activation markers on CD4+ and CD8+ T cell subsets upon T. cruzi and ribosomal protein activation. PBMC isolated from patients with chronic Chagas' disease Cardiomyopathy (CCC; n = 27) and non-infected individuals (NI; n = 20) were seeded at 2.5×10 6 cells/well and stimulated with T. cruzi lysate, P2β or CP0 proteins (10 µg/ml) or medium alone for 6 days. PBMC were stained with CD3-APC, CD4-PE-Cy5 or CD8-PE-Cy5 and activation marker-specific labeled antibodies (CD25-FITC and HLA-DR-PE) prior to flow cytometry analysis. 10,000–15,000 events in the lymphocyte gate (R1 gate) were acquired using a FACSAria flow cytometer (Becton Dickinson); dead cells were excluded by forward vs side-scatter (FSC/SSC) gating. A) Gate-pathway used to determine the activation expression in the populations graphed in B. B) Results were expressed as the percentage of CD25+ or HLA-DR+ cells in CD3+CD4+ (R2 gate) or CD3+CD8+ (R3 gate) lymphocytes. Horizontal lines represent the median and percentiles 25–75th, vertical lines represent percentiles 5–95th. Statistical analysis was performed using the Mann-Whitney U Test, *** P

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Cytokine Production but Lack of Proliferation in Peripheral Blood Mononuclear Cells from Chronic Chagas' Disease Cardiomyopathy Patients in Response to T. cruzi Ribosomal P Proteins

    doi: 10.1371/journal.pntd.0002906

    Figure Lengend Snippet: Activation markers on CD4+ and CD8+ T cell subsets upon T. cruzi and ribosomal protein activation. PBMC isolated from patients with chronic Chagas' disease Cardiomyopathy (CCC; n = 27) and non-infected individuals (NI; n = 20) were seeded at 2.5×10 6 cells/well and stimulated with T. cruzi lysate, P2β or CP0 proteins (10 µg/ml) or medium alone for 6 days. PBMC were stained with CD3-APC, CD4-PE-Cy5 or CD8-PE-Cy5 and activation marker-specific labeled antibodies (CD25-FITC and HLA-DR-PE) prior to flow cytometry analysis. 10,000–15,000 events in the lymphocyte gate (R1 gate) were acquired using a FACSAria flow cytometer (Becton Dickinson); dead cells were excluded by forward vs side-scatter (FSC/SSC) gating. A) Gate-pathway used to determine the activation expression in the populations graphed in B. B) Results were expressed as the percentage of CD25+ or HLA-DR+ cells in CD3+CD4+ (R2 gate) or CD3+CD8+ (R3 gate) lymphocytes. Horizontal lines represent the median and percentiles 25–75th, vertical lines represent percentiles 5–95th. Statistical analysis was performed using the Mann-Whitney U Test, *** P

    Article Snippet: In all cases, 10,000 to 15,000 events in the lymphocyte gate were acquired using a FACSAria flow cytometer (Becton Dickinson).

    Techniques: Activation Assay, Isolation, Countercurrent Chromatography, Infection, Staining, Marker, Labeling, Flow Cytometry, Cytometry, Expressing, MANN-WHITNEY