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  • el4  (ATCC)
    85
    ATCC el4
    Adoptive Transfer of a Limited Number of gp33-Specific CD8 + T Cells Does Not Significantly Alter the Overall Response to gp33, gp276, or np396 during the Immune Response to LCMV Primary CTL activity of splenocytes from uninfected B6.PL mice that received 10 5 P14 cells (open circles) and day 8 LCMV-infected mice (filled symbols) that received no P14 cells (squares), 10 3 P14 cells (diamonds), 10 5 P14 cells (triangles), or 10 6 P14 cells (crosses) 1 day prior to infection. Lysis was determined on day 8 of infection by 51 Cr-release assay using <t>EL4</t> targets without additional peptide (A) and EL4 pulsed with gp33 (B), gp276 (C), or np396 (D) peptides. Data are presented as the average specific lysis of targets by effectors from three individual mice per group, with standard deviation indicated by error bars (some of which fall within the symbols). Two repetitions of the experiment yielded similar results.
    El4, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 419 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher brdu
    Histopathologic detection of engrafted MSCs. Prussian blue staining (A) and <t>immunohistochemistry</t> <t>anti-BrdU</t> (B) at early time points (1 day) demonstrate single cells found within the capillaries (arrow heads) throughout the ipsilateral brain hemisphere.
    Brdu, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 4708 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore rat c6 bu 1 glioma cells
    Histopathologic detection of engrafted MSCs. Prussian blue staining (A) and <t>immunohistochemistry</t> <t>anti-BrdU</t> (B) at early time points (1 day) demonstrate single cells found within the capillaries (arrow heads) throughout the ipsilateral brain hemisphere.
    Rat C6 Bu 1 Glioma Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    SouthernBiotech bu1
    Histopathologic detection of engrafted MSCs. Prussian blue staining (A) and <t>immunohistochemistry</t> <t>anti-BrdU</t> (B) at early time points (1 day) demonstrate single cells found within the capillaries (arrow heads) throughout the ipsilateral brain hemisphere.
    Bu1, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher monoclonal antibody bu 1
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Monoclonal Antibody Bu 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    SouthernBiotech anti bu 1 fitc
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Anti Bu 1 Fitc, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher bu1 75
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Bu1 75, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Accurate Chemical & Scientific Corporation bu1 75
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Bu1 75, supplied by Accurate Chemical & Scientific Corporation, used in various techniques. Bioz Stars score: 89/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    AbCys s a bu1 75
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Bu1 75, supplied by AbCys s a, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    GE Healthcare mouse monoclonal antibody bu 1
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Mouse Monoclonal Antibody Bu 1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 80/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare bu1 75
    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) <t>Immunohistochemical</t> detection of <t>BrdU</t> incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.
    Bu1 75, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Cambridge Bioscience phycoerythrin labeled anti bu 1
    Characterization of B- and T-cell responses for intact, surgically bursectomized, or cyclophosphamide-treated chickens after infection with serovar Typhimurium. Enzyme-linked immunosorbent assay for STAgP-specific IgM, IgG, and IgA (A), percentage of <t>BU-1</t> + cells in the spleen (B), proliferation of splenocytes (48 dpi) in response to STAgP (C). Each error bar represents the standard error of the mean, and an asterisk indicates a significant difference from the intact controls (A and B) and between antigen-stimulated and unstimulated controls (C) using Student's t test ( P
    Phycoerythrin Labeled Anti Bu 1, supplied by Cambridge Bioscience, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Bio-Rad bu1 75 icr1
    Characterization of B- and T-cell responses for intact, surgically bursectomized, or cyclophosphamide-treated chickens after infection with serovar Typhimurium. Enzyme-linked immunosorbent assay for STAgP-specific IgM, IgG, and IgA (A), percentage of <t>BU-1</t> + cells in the spleen (B), proliferation of splenocytes (48 dpi) in response to STAgP (C). Each error bar represents the standard error of the mean, and an asterisk indicates a significant difference from the intact controls (A and B) and between antigen-stimulated and unstimulated controls (C) using Student's t test ( P
    Bu1 75 Icr1, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Bio-Rad anti bu 1
    Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and <t>Bu-1</t> + B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1 + B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1 + B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1 + B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.
    Anti Bu 1, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    SouthernBiotech bu 1 antibody
    Quantitation of lymphocyte phenotypes in non-PALT compartments . Means (+sem) of the number of positive cells/area for each phenotype at the time points sampled. <t>Bu-1</t> + lymphocytes were not detectable in this compartment. Note differences in the scale compared to Figure 6.
    Bu 1 Antibody, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam bu1 75
    The detection of EdU after the click reaction with various fluorescent azido dyes. A. The results of the detection of EdU using the antibody clone <t>BU1/75</t> and anti-rat antibody conjugated with FITC (F) or Cy3 (C) after the click reaction with 0.2 mM fluorescent azido-dyes or without the click reaction (control) in fixed and permeabilised cells labelled for 10 minutes with EdU are shown. The cells were incubated with 4N HCl prior to the click reaction. The images were acquired for various times in order to demonstrate that the EdU-derived signal of the anti-BrdU antibody is not completely removed by the click reaction with an elevated concentration of azido-dyes. The images in the inserts were acquired for the same time (190 ms for cells labelled with anti-rat antibody FITC conjugate and 74 ms for cells labelled with anti-rat Cy3 conjugate). Their intensity therefore reflects the decrease of the EdU-derived signal of the anti-BrdU antibody signal after the click reaction with respect to the control cells. Barr: 20 µm. B. The level of the suppression of the signal is shown for the individual azido dyes as the ratio (R) between the time length necessary to achieve the first signs of saturation in the image of the evaluated sample and in the image of control sample without the click reaction (light grey columns). Alternatively, we used the ratio between the mean intensity of the images of nuclei of the control cells and sample cells (dark grey columns).
    Bu1 75, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    GE Healthcare bu1
    Analysis of the interaction of PGL2 and <t>BU1</t> with APG. A) Interaction between PGL2 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of Trx-PGL2. Proteins co-precipitated with the amylose resin were detected by immunoblotting using anti-His antibody (upper). B) Interaction between BU1 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of GST-BU1. No co-precipitation was detected by immunoblotting using anti-GST antibody (upper). Ponceau staining indicated comparative amounts of MBP (●) and MBP-APG (◆).
    Bu1, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Adoptive Transfer of a Limited Number of gp33-Specific CD8 + T Cells Does Not Significantly Alter the Overall Response to gp33, gp276, or np396 during the Immune Response to LCMV Primary CTL activity of splenocytes from uninfected B6.PL mice that received 10 5 P14 cells (open circles) and day 8 LCMV-infected mice (filled symbols) that received no P14 cells (squares), 10 3 P14 cells (diamonds), 10 5 P14 cells (triangles), or 10 6 P14 cells (crosses) 1 day prior to infection. Lysis was determined on day 8 of infection by 51 Cr-release assay using EL4 targets without additional peptide (A) and EL4 pulsed with gp33 (B), gp276 (C), or np396 (D) peptides. Data are presented as the average specific lysis of targets by effectors from three individual mice per group, with standard deviation indicated by error bars (some of which fall within the symbols). Two repetitions of the experiment yielded similar results.

    Journal: Immunity

    Article Title: Massive Expansion of Antigen-Specific CD8+ T Cells during an Acute Virus Infection

    doi:

    Figure Lengend Snippet: Adoptive Transfer of a Limited Number of gp33-Specific CD8 + T Cells Does Not Significantly Alter the Overall Response to gp33, gp276, or np396 during the Immune Response to LCMV Primary CTL activity of splenocytes from uninfected B6.PL mice that received 10 5 P14 cells (open circles) and day 8 LCMV-infected mice (filled symbols) that received no P14 cells (squares), 10 3 P14 cells (diamonds), 10 5 P14 cells (triangles), or 10 6 P14 cells (crosses) 1 day prior to infection. Lysis was determined on day 8 of infection by 51 Cr-release assay using EL4 targets without additional peptide (A) and EL4 pulsed with gp33 (B), gp276 (C), or np396 (D) peptides. Data are presented as the average specific lysis of targets by effectors from three individual mice per group, with standard deviation indicated by error bars (some of which fall within the symbols). Two repetitions of the experiment yielded similar results.

    Article Snippet: EL4 (ATCC TIB-41) are a B6-derived (H-2b ), MHC class II– thymoma cell line and were maintained in RP10 (RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum, 2 mM l -glutamine, and antibiotics).

    Techniques: Adoptive Transfer Assay, CTL Assay, Activity Assay, Mouse Assay, Infection, Lysis, Release Assay, Standard Deviation

    Antigen-Specific Cytolytic Activity Is Proportional to the Frequency of IFNγ-Secreting Cells (A) Erythrocyte-depleted spleen cells were prepared from a pair of day 8 LCMV-infected B6 mice and were used to determine primary ex vivo CTL activity in an 8 hr 51 Cr-release assay against EL4 target cells uncoated or coated with the indicated LCMV peptides. (B) The same pool of effector cells was used in an ELISPOT assay to determine the frequency of cells secreting IFNγ in response to LCMV epitope peptides. The number of CD8 + cells was determined by FACS analysis. Error bars indicate the standard deviation of triplicate samples. Three additional experiments yielded similar results. d8, day 8. (C) The titration of spots per well versus number of LCMV day 8 splenocytes per well is shown for ELISPOT data from a separate experiment. Splenocytes were stimulated in triplicate samples with either control EL4 cells (open circles) or with gp33-coated EL4 cells (filled squares). Stimulation with gp276- or np396-coated EL4 cells resulted in similarly linear responses (data not shown).

    Journal: Immunity

    Article Title: Massive Expansion of Antigen-Specific CD8+ T Cells during an Acute Virus Infection

    doi:

    Figure Lengend Snippet: Antigen-Specific Cytolytic Activity Is Proportional to the Frequency of IFNγ-Secreting Cells (A) Erythrocyte-depleted spleen cells were prepared from a pair of day 8 LCMV-infected B6 mice and were used to determine primary ex vivo CTL activity in an 8 hr 51 Cr-release assay against EL4 target cells uncoated or coated with the indicated LCMV peptides. (B) The same pool of effector cells was used in an ELISPOT assay to determine the frequency of cells secreting IFNγ in response to LCMV epitope peptides. The number of CD8 + cells was determined by FACS analysis. Error bars indicate the standard deviation of triplicate samples. Three additional experiments yielded similar results. d8, day 8. (C) The titration of spots per well versus number of LCMV day 8 splenocytes per well is shown for ELISPOT data from a separate experiment. Splenocytes were stimulated in triplicate samples with either control EL4 cells (open circles) or with gp33-coated EL4 cells (filled squares). Stimulation with gp276- or np396-coated EL4 cells resulted in similarly linear responses (data not shown).

    Article Snippet: EL4 (ATCC TIB-41) are a B6-derived (H-2b ), MHC class II– thymoma cell line and were maintained in RP10 (RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum, 2 mM l -glutamine, and antibiotics).

    Techniques: Activity Assay, Infection, Mouse Assay, Ex Vivo, CTL Assay, Release Assay, Enzyme-linked Immunospot, FACS, Standard Deviation, Titration

    Both Adoptively Transferred P14 and Host CD8 + T Cells Are Functionally Active in LCMV-Infected PL/P14 Chimeras A group of three B6.PL mice received 10 4 P14 spleen cells. The next day the PL/P14 chimeras (C, F, and I) and groups of three B6 (A, D, and G) and B6.PL (B, E, and H) mice were infected with LCMV. On day 8 of infection, splenocytes pooled for each group were prepared and treated with rabbit complement alone (A–C), with anti-Thy1.1 (anti-host) plus complement (D–F), or with anti-Thy1.2 (anti-donor) plus complement (G–I), and the ability of the surviving cells to lyse uncoated EL4 targets and EL4 cells coated with the indicated LCMV peptides was determined in a 51 Cr-release assay.

    Journal: Immunity

    Article Title: Massive Expansion of Antigen-Specific CD8+ T Cells during an Acute Virus Infection

    doi:

    Figure Lengend Snippet: Both Adoptively Transferred P14 and Host CD8 + T Cells Are Functionally Active in LCMV-Infected PL/P14 Chimeras A group of three B6.PL mice received 10 4 P14 spleen cells. The next day the PL/P14 chimeras (C, F, and I) and groups of three B6 (A, D, and G) and B6.PL (B, E, and H) mice were infected with LCMV. On day 8 of infection, splenocytes pooled for each group were prepared and treated with rabbit complement alone (A–C), with anti-Thy1.1 (anti-host) plus complement (D–F), or with anti-Thy1.2 (anti-donor) plus complement (G–I), and the ability of the surviving cells to lyse uncoated EL4 targets and EL4 cells coated with the indicated LCMV peptides was determined in a 51 Cr-release assay.

    Article Snippet: EL4 (ATCC TIB-41) are a B6-derived (H-2b ), MHC class II– thymoma cell line and were maintained in RP10 (RPMI 1640 supplemented with 10% heat-inactivated fetal calf serum, 2 mM l -glutamine, and antibiotics).

    Techniques: Infection, Mouse Assay, Release Assay

    Histopathologic detection of engrafted MSCs. Prussian blue staining (A) and immunohistochemistry anti-BrdU (B) at early time points (1 day) demonstrate single cells found within the capillaries (arrow heads) throughout the ipsilateral brain hemisphere.

    Journal: Stroke; a journal of cerebral circulation

    Article Title: Dual-Modality Monitoring of Targeted Intraarterial Delivery of Mesenchymal Stem Cells After Transient Ischemia

    doi: 10.1161/STROKEAHA.107.502047

    Figure Lengend Snippet: Histopathologic detection of engrafted MSCs. Prussian blue staining (A) and immunohistochemistry anti-BrdU (B) at early time points (1 day) demonstrate single cells found within the capillaries (arrow heads) throughout the ipsilateral brain hemisphere.

    Article Snippet: To determine the efficiency of cell labeling as well as to correlate the histopathology with MRI, tissue preparations were processed for Prussian blue for Feridex iron and immunohistochemistry for BrdU (rat monoclonal antibody OBT0030, 1:400; Accurate Chemicals, as primary, and goat-anti rat 488, , 1:300, Molecular Probes, as secondary).

    Techniques: Staining, Immunohistochemistry

    Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) Immunohistochemical detection of BrdU incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.

    Journal: Molecular and Cellular Biology

    Article Title: ErbB2/Neu-Induced, Cyclin D1-Dependent Transformation Is Accelerated in p27-Haploinsufficient Mammary Epithelial Cells but Impaired in p27-Null Cells

    doi: 10.1128/MCB.22.7.2204-2219.2002

    Figure Lengend Snippet: Increased proliferation and decreased apoptosis in MMTV - neu/p27 +/− mammary glands; decreased proliferation in MMTV - neu-p27 −/− mammary glands. (A to C) Hematoxylin and eosin (H E) staining of sections from MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mammary glands at 60 days (A) and 120 days (B and C). (D) Immunohistochemical detection of BrdU incorporation into the mammary glands of 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 25 μm. (E) TUNEL analysis of mammary glands taken from 120-day-old MMTV - neu/p27 +/+ , MMTV - neu/p27 +/− , and MMTV - neu/p27 −/− mice. Bar, 50 μm. The average percentages of BrdU-positive and TUNEL-positive cells are indicated in the lower right corner of each panel.

    Article Snippet: Immunohistochemical detection of BrdU incorporation was performed using a monoclonal BrdU antibody (Zymed) according to the manufacturer's instructions.

    Techniques: Staining, Immunohistochemistry, BrdU Incorporation Assay, Mouse Assay, TUNEL Assay

    ErbB2 increases proliferation and anchorage-independent growth in p27 +/− cells but not in p27 −/− cells. (A) Left panel, quantification of the percentage of nuclei from cultured, infected cells that were BrdU positive [(number of BrdU-positive nuclei/total number of nuclei) × 100], shown as the means from three experiments. Error bars represent standard deviations. Black bars, p27 +/+ cells; gray bars, p27 +/− cells; white bars, p27 −/− cells. Right panels, immunohistochemical detection of BrdU-positive nuclei of pBabe- erbB2 -infected PMECs. Samples shown are representative of three independent experiments. (B) Unsynchronized cells were stained with propidium iodide and then sorted using flow cytometry to determine the proportion of cells in each phase of the cell cycle. Results are reported as the percentage of the cell population in S phase and are presented as the averages from three independent experiments. Error bars represent the standard deviations. (C) Cells were grown in soft agar for 2 weeks. The number of colonies per 35-mm-diameter plate was counted with an automated colony counter. Each sample was analyzed in triplicate, and the experiment was repeated three times. The values shown are the averages. Error bars represent the standard errors of the means.

    Journal: Molecular and Cellular Biology

    Article Title: ErbB2/Neu-Induced, Cyclin D1-Dependent Transformation Is Accelerated in p27-Haploinsufficient Mammary Epithelial Cells but Impaired in p27-Null Cells

    doi: 10.1128/MCB.22.7.2204-2219.2002

    Figure Lengend Snippet: ErbB2 increases proliferation and anchorage-independent growth in p27 +/− cells but not in p27 −/− cells. (A) Left panel, quantification of the percentage of nuclei from cultured, infected cells that were BrdU positive [(number of BrdU-positive nuclei/total number of nuclei) × 100], shown as the means from three experiments. Error bars represent standard deviations. Black bars, p27 +/+ cells; gray bars, p27 +/− cells; white bars, p27 −/− cells. Right panels, immunohistochemical detection of BrdU-positive nuclei of pBabe- erbB2 -infected PMECs. Samples shown are representative of three independent experiments. (B) Unsynchronized cells were stained with propidium iodide and then sorted using flow cytometry to determine the proportion of cells in each phase of the cell cycle. Results are reported as the percentage of the cell population in S phase and are presented as the averages from three independent experiments. Error bars represent the standard deviations. (C) Cells were grown in soft agar for 2 weeks. The number of colonies per 35-mm-diameter plate was counted with an automated colony counter. Each sample was analyzed in triplicate, and the experiment was repeated three times. The values shown are the averages. Error bars represent the standard errors of the means.

    Article Snippet: Immunohistochemical detection of BrdU incorporation was performed using a monoclonal BrdU antibody (Zymed) according to the manufacturer's instructions.

    Techniques: Cell Culture, Infection, Immunohistochemistry, Staining, Flow Cytometry, Cytometry

    Endothelial cells induce differentiation and expansion of macrophages in vitro. (A) Schematic representation of the coculture system that includes murine hematopoietic cells (HCs) and immortalized mouse endothelial cells (IMECs). All HCs express DsRed fluorescent protein. (B) Phase contrast (left) and fluorescent (right) micrographs depict the formation of DsRed + colonies (arrow) during coculture. Note that the colonies were phase-dim or invisible under phase view. Scale bar, 200 μm. (C) Photomicrographs of isolated bone marrow (BM) cells and cells from day 27 colonies (CC, arrow) after May-Grünwald-Giemsa staining. Scale bar, 50 μm. (D) FACS profile (left) indicates that the large majority of cells in the colonies were F4/80 + /Mac-1 + (n = 3). (E) Fluorescent micrographs of bromodeoxyuridine (BrdU) staining indicating active proliferation within the colony. Day 10 colonies were pulse-labeled with BrdU for 8 hours and then costained for anti-BrdU (red) and anti-F4/80 (green) antibodies. Arrow, cells positive for both F4/80 and BrdU. Arrowhead indicates F4/80 + cells not stained for BrdU. DAPI indicates staining for nucleus (blue). Scale bar, 50 μm. (F) Colony cells uptake DiI-ac-LDL. Cells were incubated with 10 μg/mL DiI-ac-LDL for 4 hours, followed by fluorescent microscopy (left). (Right) FACS profile shows the majority colony cells take up Dil-ac-LDL. CD31, to exclude possible IMEC contamination. Scale bar, 200 μm.

    Journal: Blood

    Article Title: Endothelial cells provide an instructive niche for the differentiation and functional polarization of M2-like macrophages

    doi: 10.1182/blood-2012-04-422758

    Figure Lengend Snippet: Endothelial cells induce differentiation and expansion of macrophages in vitro. (A) Schematic representation of the coculture system that includes murine hematopoietic cells (HCs) and immortalized mouse endothelial cells (IMECs). All HCs express DsRed fluorescent protein. (B) Phase contrast (left) and fluorescent (right) micrographs depict the formation of DsRed + colonies (arrow) during coculture. Note that the colonies were phase-dim or invisible under phase view. Scale bar, 200 μm. (C) Photomicrographs of isolated bone marrow (BM) cells and cells from day 27 colonies (CC, arrow) after May-Grünwald-Giemsa staining. Scale bar, 50 μm. (D) FACS profile (left) indicates that the large majority of cells in the colonies were F4/80 + /Mac-1 + (n = 3). (E) Fluorescent micrographs of bromodeoxyuridine (BrdU) staining indicating active proliferation within the colony. Day 10 colonies were pulse-labeled with BrdU for 8 hours and then costained for anti-BrdU (red) and anti-F4/80 (green) antibodies. Arrow, cells positive for both F4/80 and BrdU. Arrowhead indicates F4/80 + cells not stained for BrdU. DAPI indicates staining for nucleus (blue). Scale bar, 50 μm. (F) Colony cells uptake DiI-ac-LDL. Cells were incubated with 10 μg/mL DiI-ac-LDL for 4 hours, followed by fluorescent microscopy (left). (Right) FACS profile shows the majority colony cells take up Dil-ac-LDL. CD31, to exclude possible IMEC contamination. Scale bar, 200 μm.

    Article Snippet: Immunofluorescent labeling was performed using standard procedures with anti–mouse BrdU (Pierce, MA3-071) and anti–mouse F4/80 (Serotech; MCA497R) antibodies.

    Techniques: In Vitro, Isolation, Staining, FACS, BrdU Staining, Labeling, Incubation, Microscopy

    Characterization of B- and T-cell responses for intact, surgically bursectomized, or cyclophosphamide-treated chickens after infection with serovar Typhimurium. Enzyme-linked immunosorbent assay for STAgP-specific IgM, IgG, and IgA (A), percentage of BU-1 + cells in the spleen (B), proliferation of splenocytes (48 dpi) in response to STAgP (C). Each error bar represents the standard error of the mean, and an asterisk indicates a significant difference from the intact controls (A and B) and between antigen-stimulated and unstimulated controls (C) using Student's t test ( P

    Journal: Infection and Immunity

    Article Title: Clearance of Enteric Salmonella enterica Serovar Typhimurium in Chickens Is Independent of B-Cell Function

    doi: 10.1128/IAI.74.2.1442-1444.2006

    Figure Lengend Snippet: Characterization of B- and T-cell responses for intact, surgically bursectomized, or cyclophosphamide-treated chickens after infection with serovar Typhimurium. Enzyme-linked immunosorbent assay for STAgP-specific IgM, IgG, and IgA (A), percentage of BU-1 + cells in the spleen (B), proliferation of splenocytes (48 dpi) in response to STAgP (C). Each error bar represents the standard error of the mean, and an asterisk indicates a significant difference from the intact controls (A and B) and between antigen-stimulated and unstimulated controls (C) using Student's t test ( P

    Article Snippet: Phycoerythrin-labeled anti-BU-1 (Cambridge Bioscience, Cambridge, United Kingdom) recognizes chicken B cells ( , ) and was used for fluorescence-activated cell sorting analysis.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay

    Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1 + B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1 + B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1 + B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1 + B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.

    Journal: Development (Cambridge, England)

    Article Title: Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage

    doi: 10.1242/dev.105593

    Figure Lengend Snippet: Confocal analysis of MacRed chicken post-hatch mononuclear phagocyte populations. (A) Splenic mononuclear phagocytes (red) and Bu-1 + B-cells (green) from a 16-week-old MacRed chicken. Rings of transgene-expressing cells can clearly be seen surrounding the ellipsoid (asterisk). (B) Bursa of Fabricius from an 8-day-old MacRed chicken: Bu-1 + B cells (green) show arrangement of the B-cell follicles; mononuclear phagocytes (red) are present in the medulla (M) and interfollicular region (red arrow), but not in the cortex (C) of B-cell follicles in the bursa of Fabricius. (C) Caecal tonsil B-cell follicle from a 10-week-old MacRed chicken, showing location of mononuclear phagocytes (red) and Bu-1 + B-cells (green). Transgene-expressing cells concentrated in the medulla region (M) of the B-cell follicle are a dense network of FDC. (D) Microglial cells (red) in the cerebellum of an 8-day-old MacRed chicken showing colocalisation with CD45 staining (green). (E) Kupffer cells (red) showing colocalisation with CSF1R (green) from a 13-week-old MacRed chicken liver. (F) Lung mononuclear phagocytes (red) and Bu-1 + B-cells (green) in the interstitial tissue of the parabronchial wall from a 16-week-old MacRed chicken. The parabronchial lumen (pb) is indicated. (G) Epidermal mononuclear phagocyte cells (red) in epidermal sheet preparation from a 10-week-old MacRed chicken. (H) Breast muscle mononuclear phagocytes (red) from a 16-week-old MacRed chicken co-expressing MHCII (green). (I) Feather pulp mononuclear phagocytes from an 8-day-old MacRed chicken (red) co-stained with CD45 (green). Scale bars: 50 µm.

    Article Snippet: Primary antibodies were added: anti-CSF1R ( ); anti-MHC II [clone 2G11 ( )]; anti-chicken CD41/61 (clone 11C3, AbD Serotec); CD45 (clone LT40, SouthernBiotech); anti-Bu-1 (clone L22, AbD Serotec); anti-chicken macrophage subset marker (clone CVI-ChNL-74.2, Prionics); anti-chicken macrophage/monoctyes (clone KUL01, AbD Serotec); and anti-chicken TCR alpha/beta (clone TCR2, AbD Serotec) all diluted by 1/50-1/500 in MST-PBS and sections incubated at 4°C overnight.

    Techniques: Expressing, Staining

    F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R -mApple-expressing cells (red) in relation to: (J) Bu-1 + B-cells (green), (K) TCR αβ (Vβ1) + T-cells (green) and (L) CVI-ChNL-74.2 + macrophages (green). Scale bars in J-L: 100 µm.

    Journal: Development (Cambridge, England)

    Article Title: Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage

    doi: 10.1242/dev.105593

    Figure Lengend Snippet: F distribution of lymphoid aggregates in the MacRed chicken gut. (A-I) External views of different regions of a 1-year-old MacRed chicken showing several scattered lymphoid aggregates in the jejunum (A-C), numerous scattered lymphoid aggregates in the ileum (D-F) and a high concentration of lymphoid aggregates in the ileum Peyer's Patch. Scale bars in A-I: 500 µm. (J-L) Immunofluorescence staining of Peyer's patches showing organisation of CSF1R -mApple-expressing cells (red) in relation to: (J) Bu-1 + B-cells (green), (K) TCR αβ (Vβ1) + T-cells (green) and (L) CVI-ChNL-74.2 + macrophages (green). Scale bars in J-L: 100 µm.

    Article Snippet: Primary antibodies were added: anti-CSF1R ( ); anti-MHC II [clone 2G11 ( )]; anti-chicken CD41/61 (clone 11C3, AbD Serotec); CD45 (clone LT40, SouthernBiotech); anti-Bu-1 (clone L22, AbD Serotec); anti-chicken macrophage subset marker (clone CVI-ChNL-74.2, Prionics); anti-chicken macrophage/monoctyes (clone KUL01, AbD Serotec); and anti-chicken TCR alpha/beta (clone TCR2, AbD Serotec) all diluted by 1/50-1/500 in MST-PBS and sections incubated at 4°C overnight.

    Techniques: Concentration Assay, Immunofluorescence, Staining, Expressing

    Quantitation of lymphocyte phenotypes in non-PALT compartments . Means (+sem) of the number of positive cells/area for each phenotype at the time points sampled. Bu-1 + lymphocytes were not detectable in this compartment. Note differences in the scale compared to Figure 6.

    Journal: BMC Immunology

    Article Title: Characterization of lymphocyte subsets over a 24-hour period in Pineal-Associated Lymphoid Tissue (PALT) in the chicken

    doi: 10.1186/1471-2172-7-1

    Figure Lengend Snippet: Quantitation of lymphocyte phenotypes in non-PALT compartments . Means (+sem) of the number of positive cells/area for each phenotype at the time points sampled. Bu-1 + lymphocytes were not detectable in this compartment. Note differences in the scale compared to Figure 6.

    Article Snippet: The Bu-1 antibody (AV20 clone, SouthernBiotechnology) was used to detect B cells and cells in the monocyte lineage.

    Techniques: Quantitation Assay

    Distribution of Bu-1 + cells . PALT is seen bordering a pineal follicle stained with anti-CD3 (red) and anit-Bu-1 (green). The arrows indicate processes of microglia, which are morphologically distinct from the round lymphocytes in PALT.

    Journal: BMC Immunology

    Article Title: Characterization of lymphocyte subsets over a 24-hour period in Pineal-Associated Lymphoid Tissue (PALT) in the chicken

    doi: 10.1186/1471-2172-7-1

    Figure Lengend Snippet: Distribution of Bu-1 + cells . PALT is seen bordering a pineal follicle stained with anti-CD3 (red) and anit-Bu-1 (green). The arrows indicate processes of microglia, which are morphologically distinct from the round lymphocytes in PALT.

    Article Snippet: The Bu-1 antibody (AV20 clone, SouthernBiotechnology) was used to detect B cells and cells in the monocyte lineage.

    Techniques: Staining

    Immunohistochemical detection of Bu1+ B ( a , b ) and CD4+ ( c , d ) as well as CD8 + ( e , f ) T cells in the lamina propria of the caecum of mono- and co-inoculated birds of Exp. A ( a , c , e ) and Exp. B ( b , d , f ) (n = 6/group). pbi post bacterial inoculation; pvi post IBDV (virus) inoculation. Error bars indicate the standard deviation (SD). abc letters indicate significant differences between groups within the same experiment at the indicated time points ( P

    Journal: Gut Pathogens

    Article Title: Infectious bursal disease virus inoculation infection modifies Campylobacter jejuni–host interaction in broilers

    doi: 10.1186/s13099-018-0241-1

    Figure Lengend Snippet: Immunohistochemical detection of Bu1+ B ( a , b ) and CD4+ ( c , d ) as well as CD8 + ( e , f ) T cells in the lamina propria of the caecum of mono- and co-inoculated birds of Exp. A ( a , c , e ) and Exp. B ( b , d , f ) (n = 6/group). pbi post bacterial inoculation; pvi post IBDV (virus) inoculation. Error bars indicate the standard deviation (SD). abc letters indicate significant differences between groups within the same experiment at the indicated time points ( P

    Article Snippet: The following mouse-anti-chicken primary unlabeled antibodies were used: anti-CD4 (clone CT-4), anti-CD8β (clone EP-42) and anti-Bu1 (21-1A1) all at a concentration of 0.05 µg/ml (Southern Biotech, provided by Biozol, Eching, Germany).

    Techniques: Immunohistochemistry, Standard Deviation

    The detection of EdU after the click reaction with various fluorescent azido dyes. A. The results of the detection of EdU using the antibody clone BU1/75 and anti-rat antibody conjugated with FITC (F) or Cy3 (C) after the click reaction with 0.2 mM fluorescent azido-dyes or without the click reaction (control) in fixed and permeabilised cells labelled for 10 minutes with EdU are shown. The cells were incubated with 4N HCl prior to the click reaction. The images were acquired for various times in order to demonstrate that the EdU-derived signal of the anti-BrdU antibody is not completely removed by the click reaction with an elevated concentration of azido-dyes. The images in the inserts were acquired for the same time (190 ms for cells labelled with anti-rat antibody FITC conjugate and 74 ms for cells labelled with anti-rat Cy3 conjugate). Their intensity therefore reflects the decrease of the EdU-derived signal of the anti-BrdU antibody signal after the click reaction with respect to the control cells. Barr: 20 µm. B. The level of the suppression of the signal is shown for the individual azido dyes as the ratio (R) between the time length necessary to achieve the first signs of saturation in the image of the evaluated sample and in the image of control sample without the click reaction (light grey columns). Alternatively, we used the ratio between the mean intensity of the images of nuclei of the control cells and sample cells (dark grey columns).

    Journal: PLoS ONE

    Article Title: Most Anti-BrdU Antibodies React with 2?-Deoxy-5-Ethynyluridine -- The Method for the Effective Suppression of This Cross-Reactivity

    doi: 10.1371/journal.pone.0051679

    Figure Lengend Snippet: The detection of EdU after the click reaction with various fluorescent azido dyes. A. The results of the detection of EdU using the antibody clone BU1/75 and anti-rat antibody conjugated with FITC (F) or Cy3 (C) after the click reaction with 0.2 mM fluorescent azido-dyes or without the click reaction (control) in fixed and permeabilised cells labelled for 10 minutes with EdU are shown. The cells were incubated with 4N HCl prior to the click reaction. The images were acquired for various times in order to demonstrate that the EdU-derived signal of the anti-BrdU antibody is not completely removed by the click reaction with an elevated concentration of azido-dyes. The images in the inserts were acquired for the same time (190 ms for cells labelled with anti-rat antibody FITC conjugate and 74 ms for cells labelled with anti-rat Cy3 conjugate). Their intensity therefore reflects the decrease of the EdU-derived signal of the anti-BrdU antibody signal after the click reaction with respect to the control cells. Barr: 20 µm. B. The level of the suppression of the signal is shown for the individual azido dyes as the ratio (R) between the time length necessary to achieve the first signs of saturation in the image of the evaluated sample and in the image of control sample without the click reaction (light grey columns). Alternatively, we used the ratio between the mean intensity of the images of nuclei of the control cells and sample cells (dark grey columns).

    Article Snippet: The following monoclonal and polyclonal primary antibodies were used in the study: BMC 9318 (mouse, Roche), B44 (mouse, Becton Dickinson), Bu20a (mouse, BioLegend), BU-33 (mouse, Sigma Aldrich), BU6-4 (mouse, Genetex), BU5.1 (mouse, Millipore), MoBu-1 (mouse, Exbio) and BU1/75 (rat, Abcam), chicken polyclonal anti-BrdU antibody (Abcam) and sheep polyclonal anti-BrdU antibody (Genetex).

    Techniques: Incubation, Derivative Assay, Concentration Assay, Mass Spectrometry

    The increase of azido dye concentration and acid treatment results in a non-specific signal. The picture shows examples of the detection of incorporated EdU in DNA by means of a click reaction with 0.02 mM fluorescent dye without the HCl treatment (images labelled as control) and of detection with 0.2 mM fluorescent azido dyes and the antibody clone BU1/75 (Cy3 anti-rat and FITC anti-rat secondary antibodies) in cells treated with 4N HCl. Note that the higher concentration of fluorescent azido dyes and the subsequent treatment with 4N HCl led to a non-specific signal mainly in the nucleolus area. Barr: 20 µm.

    Journal: PLoS ONE

    Article Title: Most Anti-BrdU Antibodies React with 2?-Deoxy-5-Ethynyluridine -- The Method for the Effective Suppression of This Cross-Reactivity

    doi: 10.1371/journal.pone.0051679

    Figure Lengend Snippet: The increase of azido dye concentration and acid treatment results in a non-specific signal. The picture shows examples of the detection of incorporated EdU in DNA by means of a click reaction with 0.02 mM fluorescent dye without the HCl treatment (images labelled as control) and of detection with 0.2 mM fluorescent azido dyes and the antibody clone BU1/75 (Cy3 anti-rat and FITC anti-rat secondary antibodies) in cells treated with 4N HCl. Note that the higher concentration of fluorescent azido dyes and the subsequent treatment with 4N HCl led to a non-specific signal mainly in the nucleolus area. Barr: 20 µm.

    Article Snippet: The following monoclonal and polyclonal primary antibodies were used in the study: BMC 9318 (mouse, Roche), B44 (mouse, Becton Dickinson), Bu20a (mouse, BioLegend), BU-33 (mouse, Sigma Aldrich), BU6-4 (mouse, Genetex), BU5.1 (mouse, Millipore), MoBu-1 (mouse, Exbio) and BU1/75 (rat, Abcam), chicken polyclonal anti-BrdU antibody (Abcam) and sheep polyclonal anti-BrdU antibody (Genetex).

    Techniques: Concentration Assay

    The suppression of the EdU signal using non-fluorescent azido molecules. The suppression of the signal provided by BU1/75 antibody after a click reaction with 2 or 20 mM 2-azidoethanol, 1-azido-2,3-dihydroxypropane or azidomethylphenylsulfide in cells labelled with EdU for 20 minutes. The images were acquired for 8 ms. Barr: 20 µm.

    Journal: PLoS ONE

    Article Title: Most Anti-BrdU Antibodies React with 2?-Deoxy-5-Ethynyluridine -- The Method for the Effective Suppression of This Cross-Reactivity

    doi: 10.1371/journal.pone.0051679

    Figure Lengend Snippet: The suppression of the EdU signal using non-fluorescent azido molecules. The suppression of the signal provided by BU1/75 antibody after a click reaction with 2 or 20 mM 2-azidoethanol, 1-azido-2,3-dihydroxypropane or azidomethylphenylsulfide in cells labelled with EdU for 20 minutes. The images were acquired for 8 ms. Barr: 20 µm.

    Article Snippet: The following monoclonal and polyclonal primary antibodies were used in the study: BMC 9318 (mouse, Roche), B44 (mouse, Becton Dickinson), Bu20a (mouse, BioLegend), BU-33 (mouse, Sigma Aldrich), BU6-4 (mouse, Genetex), BU5.1 (mouse, Millipore), MoBu-1 (mouse, Exbio) and BU1/75 (rat, Abcam), chicken polyclonal anti-BrdU antibody (Abcam) and sheep polyclonal anti-BrdU antibody (Genetex).

    Techniques: Mass Spectrometry

    Analysis of the interaction of PGL2 and BU1 with APG. A) Interaction between PGL2 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of Trx-PGL2. Proteins co-precipitated with the amylose resin were detected by immunoblotting using anti-His antibody (upper). B) Interaction between BU1 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of GST-BU1. No co-precipitation was detected by immunoblotting using anti-GST antibody (upper). Ponceau staining indicated comparative amounts of MBP (●) and MBP-APG (◆).

    Journal: Breeding Science

    Article Title: An atypical bHLH protein encoded by POSITIVE REGULATOR OF GRAIN LENGTH 2 is involved in controlling grain length and weight of rice through interaction with a typical bHLH protein APG

    doi: 10.1270/jsbbs.62.133

    Figure Lengend Snippet: Analysis of the interaction of PGL2 and BU1 with APG. A) Interaction between PGL2 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of Trx-PGL2. Proteins co-precipitated with the amylose resin were detected by immunoblotting using anti-His antibody (upper). B) Interaction between BU1 and APG in vitro analyzed by pull-down assay. Amylose resin–bound MBP-APG or MBP was incubated with an equal amount of GST-BU1. No co-precipitation was detected by immunoblotting using anti-GST antibody (upper). Ponceau staining indicated comparative amounts of MBP (●) and MBP-APG (◆).

    Article Snippet: BU1 was amplified from lemma/palea cDNA (FBU1e: 5′-GGAATTCATGTCGAGCCGGAGGT CGTC-3′ and RBU1xh: 5′-CCCTCGAGCTAGCGGAGGA GGCTGCGGA-3′) and sub-cloned into pGEX-4T-1 (GE, Healthcare) at Eco RI/Xho I. MBP-APG was generated as described in .

    Techniques: In Vitro, Pull Down Assay, Incubation, Staining

    Model for regulation of grain length by bHLH proteins in rice. Typical bHLH proteins APG and a hypothetical protein, bHLHx, negatively regulate rice grain length. Atypical bHLH proteins PGL1 and PGL2 function as positive regulators of grain length and suppress the function of APG through heterodimerization, while the closest homolog of PGL2, BU1, does not. BU1 is assumed to be involved in regulation of grain length through heterodimerization with bHLHx to suppress its function. Dotted lines indicate a hypothetical pathway.

    Journal: Breeding Science

    Article Title: An atypical bHLH protein encoded by POSITIVE REGULATOR OF GRAIN LENGTH 2 is involved in controlling grain length and weight of rice through interaction with a typical bHLH protein APG

    doi: 10.1270/jsbbs.62.133

    Figure Lengend Snippet: Model for regulation of grain length by bHLH proteins in rice. Typical bHLH proteins APG and a hypothetical protein, bHLHx, negatively regulate rice grain length. Atypical bHLH proteins PGL1 and PGL2 function as positive regulators of grain length and suppress the function of APG through heterodimerization, while the closest homolog of PGL2, BU1, does not. BU1 is assumed to be involved in regulation of grain length through heterodimerization with bHLHx to suppress its function. Dotted lines indicate a hypothetical pathway.

    Article Snippet: BU1 was amplified from lemma/palea cDNA (FBU1e: 5′-GGAATTCATGTCGAGCCGGAGGT CGTC-3′ and RBU1xh: 5′-CCCTCGAGCTAGCGGAGGA GGCTGCGGA-3′) and sub-cloned into pGEX-4T-1 (GE, Healthcare) at Eco RI/Xho I. MBP-APG was generated as described in .

    Techniques: