b220 apc cy7  (Cytek Biosciences)

Journal: PLoS ONE

Article Title: BCL2A1a Over-Expression in Murine Hematopoietic Stem and Progenitor Cells Decreases Apoptosis and Results in Hematopoietic Transformation

doi: 10.1371/journal.pone.0048267

Figure Lengend Snippet: (A) Growth curves for BAF3 cells grown with or without murine IL-3. Viable cell numbers are shown over time. (B) Cell cycle analysis on BAF3 cells grown without IL-3. (C) Apoptosis assessed via Annexin V-PE staining in the absence of IL-3. All experiments were repeated 3 times and carried out in triplicates. Figure shows results obtained for one representative experiment. Data averages plus or minus standard deviations were plotted. HA-mBCL2A1a and HA-hBCL2A1 are respectively murine and human HA-BCL2A1.

Article Snippet: Peripheral blood, bone marrow, and spleen cells were resuspended in FACS buffer (2.7 mM Potassium chloride, 1.5 mM Sodium phosphate dibasic heptahydrate, 1.5 mM Potassium phosphate monobasic, Sodium chloride 137 mM, Sodium azide 7.7 mM, 1% (w/v) BSA in water) and incubated with the following cocktail of antibodies from BD Pharmingen™ (BD Bioscience, San Diego, CA): T-cells-anti-mouse CD3e (Hamster, APC), granulocytes and NK cells-anti-mouse CD11b (Rat, PE-Cy7), B-cells-anti-mouse CD45R/B220 (Rat, APC-Cy7).

Techniques: Cell Cycle Assay, Staining

Journal: PLoS ONE

Article Title: BCL2A1a Over-Expression in Murine Hematopoietic Stem and Progenitor Cells Decreases Apoptosis and Results in Hematopoietic Transformation

doi: 10.1371/journal.pone.0048267

Figure Lengend Snippet: (A) Micrographs obtained from H&E stained slides of sternal sections or cytospins from bone marrow cells. Primary BCL2A1a mice #25 and #33 as well as secondary mouse #24-12 are represented. Sternal sections were appreciated at 20X magnification whereas blastic cells from cytospins were appreciated at 100X magnification. (B) Example of immunohistochemical staining of lymph nodes from primary BCL2A1a mouse (#33). Lymph node sections were stained with antibodies directed against antigens including B220 for B-cells, CD3 for T-cells, TdT for immature leukemic cells, lysozyme for myeloid cells, CD68 for monocytes or histiocytes. Sections were also stained with Hematoxylin and Eosin (H&E).

Article Snippet: Peripheral blood, bone marrow, and spleen cells were resuspended in FACS buffer (2.7 mM Potassium chloride, 1.5 mM Sodium phosphate dibasic heptahydrate, 1.5 mM Potassium phosphate monobasic, Sodium chloride 137 mM, Sodium azide 7.7 mM, 1% (w/v) BSA in water) and incubated with the following cocktail of antibodies from BD Pharmingen™ (BD Bioscience, San Diego, CA): T-cells-anti-mouse CD3e (Hamster, APC), granulocytes and NK cells-anti-mouse CD11b (Rat, PE-Cy7), B-cells-anti-mouse CD45R/B220 (Rat, APC-Cy7).

Techniques: Staining, Immunohistochemical staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Proportion of main immune cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( A–D ) Proportion of B220 + cells, CD3 + cells, CD11c + cells, CD49b + cells, and CD11b + cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( E–H ) Proportion of subpopulations of B cells in peripheral organs of CJ mice, Bmal1 -/- mice, and Per1 -/- Per2 -/- mice. ( I ) Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells, MFI of IL10 and IL33 on B220 + CD1d + CD5 + cells in IELs from DSS-treated or untreated Bmal1 -/- and WT mice. ( J ) Proportion of CD1d + CD5 + cells of B220 + cells in IELs. ( K ) MFI of IL33 and ( L ) IL10 on B220 + CD1d + CD5 + in IELs. ( M ) Rhythmic oscillation of proportion of B220 + CD1d + CD5 + cells in the IELs and ( N ) hepatic lymphocytes from DSS-treated Bmal1 -/- and WT mice within 24 hours. The P value is estimated by JTK cycle analysis. FSC, forward scatter; NC, non-specific control; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: The proportion of Breg cells expressing PDL1 in IELs, SPLs, and PBLs in DSS-treated and untreated Bmal1 -/- mice. ( A ) Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells and MFI of PDL1 on B220 + CD1d + CD5 + cells in IELs, ( C ) SPLs, and ( E ) PBLs of DSS-treated and untreated Bmal1 -/- and WT mice. ( B ) MFI of PDL1 on B220 + CD1d + CD5 + cells and proportion of B220 + PDL1 + cells, CD5 + PDL1 + cells, and CD1d + PDL1 + cells from CD45 + cells in IELs of DSS-treated and untreated Bmal1 -/- and WT mice. MFI of PDL1 on B220 + CD1d + CD5 + cells in ( D ) SPLs and ( F ) PBLs of DSS-treated and untreated Bmal1 -/- and WT mice. APC, Allophycocyanin; FITC, Fluoresceine Isothiocyanate; FSC, forward scatter; NC, non-specific control; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Expressing, Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Effects of adoptive cell transfer treatment using B cells isolated from WT mice in DSS-treated Bmal1 -/- mice or using B cells from Bmal1 -/- mice in DSS-treated WT mice. ( A ) Gross appearance of colorectum, ( B ) body weight, ( C ) colon length, ( D ) histologic score of colon, and ( E ) H&E of colon from DSS-treated Bmal1 -/- mice, Bmal1 -/- mice receiving B cells isolated from Bmal1 -/- mice, and from WT mice, and WT mice receiving B cells isolated from WT mice. ( F ) Proportion of CD1d + CD5 + cells gated from B220 + cells in IELs. ( G ) MFI of IL10 and ( H ) PDL1 on B220 + CD1d + CD5 + in IELs. ( I ) Fluorescence-activated cell sorter (FACS) graphs of B220 + CD1d + CD5 + cells, and MFI of PDL1 and IL10 on B220 + CD1d + CD5 + cells in IELs. ( J–L ) Proportion of CD1d + CD5 + cells gated from B220 + cells, MFI of IL10 and PDL1 on B220 + CD1d + CD5 + cells in SPLs. ( M–O ) Proportion of CD1d + CD5 + cells from B220 + cells, MFI of IL10 and PDL1 on B220 + CD1d + CD5 + cells in PBLs. ( P ) Gross appearance of colorectum, ( Q ) body weight, ( R ) colon length, ( S ) H&E of colon, and ( T ) histologic score of colon from WT mice and WT mice receiving B cells from Bmal1 -/- mice. ( U ) FACS graphs showing B220 + CD1d + CD5 + cells, and MFI of IL10 and PDL1 on B220 + CD1d + CD5 + cells in IELs. ∗ P < .05. APC, Allophycocyanin; FITC, Fluoresceine Isothiocyanate; FSC, forward scatter; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Isolation, Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells, and the MFI of PDL1 and IL10 on ( A ) B220 + CD1d + CD5 + cells in SPLs and ( B ) PBLs from DSS-treating Bmal1 -/- mice, Bmal1 -/- receiving B cells from Bmal1 -/- mice, Bmal1 -/- receiving B cells from WT mice, and WT receiving B cells from WT mice. FACS graphs showing B220 + CD1d + CD5 + cells, and MFI of PDL1 and IL10 on B220 + CD1d + CD5 + cells in ( C ) SPLs and ( D ) PBLs from DSS-treating WT mice and WT receiving B cells from Bmal1 -/- mice. APC, Allophycocyanin; FITC, Fluoresceine Isothiocyanate; FSC, forward scatter; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Effects of adoptive cell transfer treatment using adoptive B cells activated in vitro and isolated from WT mice in DSS-untreated Bmal1 -/- mice or using B cells from Bmal1 -/- mice in DSS-untreated WT mice. ( A ) Gross appearance of colorectum, ( B ) body weight, ( C ) colon length, ( D ) histologic score of colon, ( E ) H&E of colon from DSS-treated Bmal1 -/- mice, Bmal1 -/- mice receiving B cells isolated from Bmal1 -/- mice and from WT mice, and WT mice receiving B cells isolated from WT mice. ( F ) Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells, and MFI of IL10 and PDL1 on B220 + CD1d + CD5 + in IELs from models. ( G ) Proportion of CD1d + CD5 + cells from B220 + cells, and MFI of ( H ) PDL1 and ( I ) IL10 on B220 + CD1d + CD5 + cells in IELs from models. ( J ) Proportion of CD1d + CD5 + cells from B220 + in SPLs, and ( K ) MFI of PDL1 and ( L ) IL10 on B220 + CD1d + CD5 + cells in SPLs from models. ( M ) Proportion of CD1d + CD5 + cells from B220 + in PBLs, and ( N ) MFI of PDL1 and ( O ) IL10 on B220 + CD1d + CD5 + cells in SPLs from models. ∗ P < .05.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: In Vitro, Isolation, Fluorescence

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: The effects of Breg cells expressing PDL1 in IELs in DSS-treated Bmal1 -/- mice. ( A ) Gross appearance of colorectum, ( B ) body weight of mice, ( C ) H&E of colon, ( D ) colon length, ( E ) histologic score of colon from DSS-treated Bmal1 -/- mice, Bmal1 -/- mice receiving B cells donated from WT mice, and Bmal1 -/- mice receiving B cells donated from WT mice and PDL1 antagonist antibody. ( F ) Fluorescence-activated cell sorter (FACS) graphs showing B220 + CD1d + CD5 + cells, and MFI of PDL1 and IL10 on B220 + CD1d + CD5 + in IELs, ( H ) SPLs, and ( J ) PBLs. ( G ) Proportion of CD1d + CD5 + gated on B220 + cells, and MFI of IL10 and PDL1 on B220 + CD1d + CD5 + cells in IELs, ( I ) SPLs, and ( K ) PBLs. ( L ) FACS graphs showing PDL1 + B220 + cells in IELs, SPLs, and PBLs of donor mice that antagonize PDL1. ( M ) Transcription levels of PDL1 in the IELs and ( N ) hepatic lymphocytes of Bmal1 -/- and WT mice among 24 hours. The P value is estimated by JTK cycle analysis. ( O ) The effects of Bmal1 and clock complexes on luciferase activities of PDL1 promoter reporter. ∗ P < .05. APC, Allophycocyanin; FITC, Fluoresceine Isothiocyanate; FSC, forward scatter; PE, Phycoerythrin; SSC, side scatter.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Expressing, Fluorescence, Luciferase

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Cell death of activated CD4 + T cells in DSS-treated Bmal1 -/- mice and the effects of PDL1 antagonist antibody. ( A ) Fluorescence-activated cell sorter (FACS) graphs and ( B ) proportions of CD8 + and ( C ) CD4 + T cells in the colon of DSS-treated Bmal1 -/- mice and WT mice. ( D ) Protein expression of Bmal1, activated caspase 3, Bcl-2 Associated X Protein, and Bcl2 in CD8 + T cells and ( E ) CD4 + T cells in the colons of DSS-treated Bmal1 -/- mice and WT mice. ( F ) FACS graphs of CD4 + Ki67 + cells, ( G ) proportions of CD4 + T cells, ( H ) CD4 + Ki67 + cells, and ( I ) CD8 + T cells in the IELs of Bmal1 -/- mice, Bmal1 -/- mice receiving B cells from WT mice, Bmal1 -/- mice receiving B cells from WT mice and PDL1 antagonist antibodies. ( J ) FACS graphs and proportion of ( K ) Annexin-v + Comp-7-AAD - B220 + or ( L ) Annexin-v + Comp-7-AAD + B220 + cells, ( M ) Annexin-v + Comp-7-AAD - CD4 + or ( N ) Annexin-v + Comp-7-AAD + CD4 + cells, and ( O ) Annexin-v + Comp-7-AAD - CD8 + or ( P ) Annexin-v + Comp-7-AAD + CD8 + cells in IELs from DSS-treated WT mice and Bmal1 -/- mice. ( Q ) FACS graphs and ( U ) proportions of CD4 + IL17 + cells in IELs. ( R ) FACS graphs and ( V ) proportions of CD4 + Foxp3 + cells in IELs. ( S ) Transcription expression of Bmal1, IL17, RORγt, IL21, and IL6 in IELs of DSS-treated WT and Bmal1 -/- mice. ( T ) Transcription expression of Forkhead Box P3, IL2, IL10, TGFβ, and IFNγ in IELs of DSS-treated WT and Bmal1 -/- mice. ∗ P < .05. APC, Allophycocyanin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Fluorescence, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: The effects of IL33 in intestinal microenvironment on PDL1 + Breg cells in DSS-treated Bmal1 -/- mice. ( A ) Transcription levels of IL4, IL6, IL10, IL21, IL33, TGFβ, and MCP-1 in the IELs of DSS-treated and untreated Bmal1 -/- and WT mice among 24 hours. The P value was estimated by JTK cycle analysis. ( B ) Transcription levels of inflammatory cytokines IL4, IL6, IL10, IL21, IL33, TGFβ, and MCP-1 in the IELs of DSS-treated Bmal1 -/- and WT mice at the same time. ( C ) Protein expressions of Bmal1 and IL33, ( D ) proportion of B220 + IL33 + cells in IELs of DSS-treated Bmal1 -/- and WT mice. ( E ) Gross appearance of colorectum, ( F ) body weight during DSS induction, ( G ) colorectal length, ( H ) histologic score of colon, ( I ) H&E of colon from DSS-treated WT mice, WT mice with IL33 antagonist antibody, Bmal1 -/- mice, and Bmal1 -/- mice with IL33 antagonist antibody. ( J ) FACS graphs and ( K ) proportion of CD5 + CD1d + cells from B220 + cells in IELs. ( L ) Fluorescence-activated cell sorter (FACS) graphs and ( M ) proportion of B220 + PDL1 + , ( N ) CD1d + PDL1 + , and ( O ) CD5 + PDL1 + cells in IELs. ( P ) PDL1 expressions in B cells isolated from WT and Bmal1 -/- spleen treated with IL21, TGFβ, or IL33 neutralizing antibodies by Western blot. ( Q ) PDL1 expression in B cells isolated from WT and Bmal1 -/- spleen treated with IL33 by Western blot. ( R ) The effects of Bmal1 and clock complexes on luciferase activities of IL33 promoter reporter with or without E-box site mutation. ( S ) ChIP assay to detect the binding of IL33 gene promoter with Bmal1 in SPLs and B cells. ∗ P < .05. GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Fluorescence, Isolation, Western Blot, Expressing, Luciferase, Mutagenesis, Binding Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Effects of disorders of biological rhythm on colitis-associated CRC and expression and prognosis of Bmal1 in clinical samples of CRC. ( A ) Gross appearance of colorectum, ( B ) H&E of colon, and ( C ) tumor numbers from CJ, Bmal1 -/- , Per1 -/- Per2 -/- , and WT mice of enteritis-related bowel cancer model. ( D ) Proportion of CD1d + CD5 + cells from B220 + cells, ( E ) IL10 + B220 + cells, ( F ) B220 + /CD1d + /CD5 + PDL1 + cells in IELs of models. ( G ) Proportion of CD8 + cells, ( H ) PD1 + CD8 + cells, ( I ) CD8 + GrazB + cells and ( J ) Fluorescence-activated cell sorter (FACS) graphs of CD8 + GrazB + in IELs of models. ( K ) Proportions and ( L ) FACS graphs of CD8 + IFNγ + cells in IELs of models. ( M ) Proportion of CD4 + T cells in IELs of models. ( N ) Proportion and ( O ) FACS graphs of CD4 + PD1 + cells in IELs of models. ( P ) Proportion of GranzB + CD4 + cells, ( Q ) IFNγ + CD4 + cells in the IELs of models. ( R ) Semiquantitative analysis and ( S ) immunochemical staining of Bmal1 + immune cells from CT, the front of the IM, and paracancerous area (Norm) in 91 CRC patients. ( T ) Survival curves of CRC patients with low or high expression of Bmal1 in the 3 regions of tumor samples. ( U ) Correlation of CD19 + cells and Bmal1 + immune cells in the 3 regions of tumor samples. ∗ P < .05.

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques: Expressing, Fluorescence, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Circadian Clock Disruption Suppresses PDL1 + Intraepithelial B Cells in Experimental Colitis and Colitis-Associated Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.02.008

Figure Lengend Snippet: Antibodies Used for Flow Cytometry

Article Snippet: APC-Cy7 anti-mouse B220 antibody , BD Pharmingen San Diego, CA , 552094 , RA3-6B2.

Techniques:

Journal: Journal of Parasitology Research

Article Title: Acute Disruption of Bone Marrow B Lymphopoiesis and Apoptosis of Transitional and Marginal Zone B Cells in the Spleen following a Blood-Stage Plasmodium chabaudi Infection in Mice

doi: 10.1155/2011/534697

Figure Lengend Snippet: Differentiation antigen phenotypes of developing and mature B2 B cells.

Article Snippet: The following antibodies were added to 100 μ L aliquots of 10 6 Fc-blocked leukocytes prepared as described above: 0.5 μ g anti-CD23-FITC (clone B3B4), 0.5 μ g anti-IL7r α -FITC (clone A7R34), 0.5 μ g anti-CD11b-FITC (clone M1/70; 0.5 mg/mL), 0.5 μ g anti-CD45R (B220)-FITC (clone RA3-6B2), 0.2 μ g anti-IgM-PE (clone II/41), 0.2 μ g anti-CD93–PE (clone AA4.1), 0.5 μ g anti-CD95-FITC (clone Jo2), 0.25 μ g hamster IgG2, κ isotype control (clone B81-3), 0.2 μ g anti-IgM PE-Cy7 (clone II/41), 0.2 μ g of anti-CD45R (B220)-PE-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD93-APC (clone AA4.1), and 0.2 μ g of anti-CD117 (ckit)-APC (clone 2B8), purchased from eBioscience (San Diego, CA); 0.2 μ g anti-CD1d-PE (clone 1b1), 0.2 μ g of anti-CD43-PE (clone 1B11), 0.2 μ g of anti-CD45R (B220)-APC-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD19-APC-Cy7 (clone 1D3), 0.2 μ g of streptavidin-PerCP, and 0.2 μ g of streptavidin-PE-Texas Red, purchased from BD Biosciences (Erembodegem, Belgium); 2 μ g of each of the following antibodies: CD3 ε , CD11b (Mac-1), Gr-1 (Ly-6G and Ly-6C) and Ter-119 (Ly-76) from the Biotin-conjugated Mouse Lineage Panel (BD Biosciences, Erembodegem, Belgium).

Techniques: Marker

Journal: Journal of Parasitology Research

Article Title: Acute Disruption of Bone Marrow B Lymphopoiesis and Apoptosis of Transitional and Marginal Zone B Cells in the Spleen following a Blood-Stage Plasmodium chabaudi Infection in Mice

doi: 10.1155/2011/534697

Figure Lengend Snippet: Extramedullary B lymphopoiesis in spleen during Pc AS infection. Spleens cells from noninfected mice and mice infected with Pc AS for 10–41 days were stained for surface markers used to define developing B cells. The total B cell precursor population, including pre-pro-B, pro-B, pre-B and immature B, was detected as 7AAD − Lin − B220 int AA4.1 hi using FACS. Data are representative of two separate experiments.

Article Snippet: The following antibodies were added to 100 μ L aliquots of 10 6 Fc-blocked leukocytes prepared as described above: 0.5 μ g anti-CD23-FITC (clone B3B4), 0.5 μ g anti-IL7r α -FITC (clone A7R34), 0.5 μ g anti-CD11b-FITC (clone M1/70; 0.5 mg/mL), 0.5 μ g anti-CD45R (B220)-FITC (clone RA3-6B2), 0.2 μ g anti-IgM-PE (clone II/41), 0.2 μ g anti-CD93–PE (clone AA4.1), 0.5 μ g anti-CD95-FITC (clone Jo2), 0.25 μ g hamster IgG2, κ isotype control (clone B81-3), 0.2 μ g anti-IgM PE-Cy7 (clone II/41), 0.2 μ g of anti-CD45R (B220)-PE-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD93-APC (clone AA4.1), and 0.2 μ g of anti-CD117 (ckit)-APC (clone 2B8), purchased from eBioscience (San Diego, CA); 0.2 μ g anti-CD1d-PE (clone 1b1), 0.2 μ g of anti-CD43-PE (clone 1B11), 0.2 μ g of anti-CD45R (B220)-APC-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD19-APC-Cy7 (clone 1D3), 0.2 μ g of streptavidin-PerCP, and 0.2 μ g of streptavidin-PE-Texas Red, purchased from BD Biosciences (Erembodegem, Belgium); 2 μ g of each of the following antibodies: CD3 ε , CD11b (Mac-1), Gr-1 (Ly-6G and Ly-6C) and Ter-119 (Ly-76) from the Biotin-conjugated Mouse Lineage Panel (BD Biosciences, Erembodegem, Belgium).

Techniques: Infection, Staining

Journal: Journal of Parasitology Research

Article Title: Acute Disruption of Bone Marrow B Lymphopoiesis and Apoptosis of Transitional and Marginal Zone B Cells in the Spleen following a Blood-Stage Plasmodium chabaudi Infection in Mice

doi: 10.1155/2011/534697

Figure Lengend Snippet: Depletion of transitional type 2 B cells in spleen during Pc AS infection. (a) Spleens cells from noninfected mice and mice infected with Pc AS for 10–41 days were stained for surface markers commonly used to define transitional T2 B cells and analyzed using FACS. Data are represented as mean of three mice per group ± SEM, two independent repeat experiments were performed and statistics are compared to uninfected controls (**) P < .01, (***) P < .001. (b), (c), and (d) Transitional T2 B cells were detected as (AA4.1 + B220 + ) IgM + CD23 + in uninfected mice and mice on day 10 and 30 pi.

Article Snippet: The following antibodies were added to 100 μ L aliquots of 10 6 Fc-blocked leukocytes prepared as described above: 0.5 μ g anti-CD23-FITC (clone B3B4), 0.5 μ g anti-IL7r α -FITC (clone A7R34), 0.5 μ g anti-CD11b-FITC (clone M1/70; 0.5 mg/mL), 0.5 μ g anti-CD45R (B220)-FITC (clone RA3-6B2), 0.2 μ g anti-IgM-PE (clone II/41), 0.2 μ g anti-CD93–PE (clone AA4.1), 0.5 μ g anti-CD95-FITC (clone Jo2), 0.25 μ g hamster IgG2, κ isotype control (clone B81-3), 0.2 μ g anti-IgM PE-Cy7 (clone II/41), 0.2 μ g of anti-CD45R (B220)-PE-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD93-APC (clone AA4.1), and 0.2 μ g of anti-CD117 (ckit)-APC (clone 2B8), purchased from eBioscience (San Diego, CA); 0.2 μ g anti-CD1d-PE (clone 1b1), 0.2 μ g of anti-CD43-PE (clone 1B11), 0.2 μ g of anti-CD45R (B220)-APC-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD19-APC-Cy7 (clone 1D3), 0.2 μ g of streptavidin-PerCP, and 0.2 μ g of streptavidin-PE-Texas Red, purchased from BD Biosciences (Erembodegem, Belgium); 2 μ g of each of the following antibodies: CD3 ε , CD11b (Mac-1), Gr-1 (Ly-6G and Ly-6C) and Ter-119 (Ly-76) from the Biotin-conjugated Mouse Lineage Panel (BD Biosciences, Erembodegem, Belgium).

Techniques: Infection, Staining

Journal: Journal of Parasitology Research

Article Title: Acute Disruption of Bone Marrow B Lymphopoiesis and Apoptosis of Transitional and Marginal Zone B Cells in the Spleen following a Blood-Stage Plasmodium chabaudi Infection in Mice

doi: 10.1155/2011/534697

Figure Lengend Snippet: Depletion of MZB but not FoB cells in spleen during Pc AS infection. (a) and (b) Spleens cells from noninfected mice and mice infected with Pc AS for 10–41 days were stained for surface markers commonly used to define MZB (a) and FoB (b) cells and analyzed using FACS. Data are represented as mean of three mice per group ± SEM, two independent repeat experiments were performed and statistics are compared to uninfected controls (**) P < .01. (c), (d), and (e) MZB and FoB cells were detected as (AA4.1 − ) B220 + CD1d + (CD23 lo CD21 hi ), respectively, (AA4.1 − ) B220 + CD1d − (CD23 hi CD21 lo ), in noninfected mice and mice on day 10 and 30 pi.

Article Snippet: The following antibodies were added to 100 μ L aliquots of 10 6 Fc-blocked leukocytes prepared as described above: 0.5 μ g anti-CD23-FITC (clone B3B4), 0.5 μ g anti-IL7r α -FITC (clone A7R34), 0.5 μ g anti-CD11b-FITC (clone M1/70; 0.5 mg/mL), 0.5 μ g anti-CD45R (B220)-FITC (clone RA3-6B2), 0.2 μ g anti-IgM-PE (clone II/41), 0.2 μ g anti-CD93–PE (clone AA4.1), 0.5 μ g anti-CD95-FITC (clone Jo2), 0.25 μ g hamster IgG2, κ isotype control (clone B81-3), 0.2 μ g anti-IgM PE-Cy7 (clone II/41), 0.2 μ g of anti-CD45R (B220)-PE-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD93-APC (clone AA4.1), and 0.2 μ g of anti-CD117 (ckit)-APC (clone 2B8), purchased from eBioscience (San Diego, CA); 0.2 μ g anti-CD1d-PE (clone 1b1), 0.2 μ g of anti-CD43-PE (clone 1B11), 0.2 μ g of anti-CD45R (B220)-APC-Cy7 (clone RA3-6B2), 0.2 μ g of anti-CD19-APC-Cy7 (clone 1D3), 0.2 μ g of streptavidin-PerCP, and 0.2 μ g of streptavidin-PE-Texas Red, purchased from BD Biosciences (Erembodegem, Belgium); 2 μ g of each of the following antibodies: CD3 ε , CD11b (Mac-1), Gr-1 (Ly-6G and Ly-6C) and Ter-119 (Ly-76) from the Biotin-conjugated Mouse Lineage Panel (BD Biosciences, Erembodegem, Belgium).

Techniques: Infection, Staining