mouse anti brdu monoclonal antibody  (Millipore)

 
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    Name:
    Monoclonal Anti 5 bromodeoxyuridine antibody
    Description:
    The antibody MoBu 1 reacts specifically with BrdU incorporated into DNA during S phase of a cell cycle The antibody MoBu 1 is also useful for detecting proliferating cells by flow cytometry or immunofluorescence staining It reacts also specifically with 5 bromouridine BrU
    Catalog Number:
    SAB4700630
    Price:
    None
    Applications:
    Monoclonal Anti-5-bromodeoxyuridine antibody produced in mouse has been used in Immunocytochemistry. The reagent is designed for Flow Cytometry analysis. Suggested working dilution for Flow Cytometry is 1-2 μg/mL of sample. Indicated dilution is recommended starting point for use of this product. Working concentrations should be determined by the investigator.
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    Structured Review

    Millipore mouse anti brdu monoclonal antibody
    Monoclonal Anti 5 bromodeoxyuridine antibody
    The antibody MoBu 1 reacts specifically with BrdU incorporated into DNA during S phase of a cell cycle The antibody MoBu 1 is also useful for detecting proliferating cells by flow cytometry or immunofluorescence staining It reacts also specifically with 5 bromouridine BrU
    https://www.bioz.com/result/mouse anti brdu monoclonal antibody/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti brdu monoclonal antibody - by Bioz Stars, 2021-04
    93/100 stars

    Images

    1) Product Images from "A chemical method for fast and sensitive detection of DNA synthesis in vivo"

    Article Title: A chemical method for fast and sensitive detection of DNA synthesis in vivo

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0712168105

    Reproducibility of EdU labeling ( A ) and comparison with BrdU ( B ). ( A ) NIH 3T3 cells labeled by incubation overnight with 2 μM EdU were fixed and reacted successively with 10 μM Alexa488-azide and 10 μM Alexa594-azide, respectively, as described in Materials and Methods . The cells were imaged by fluorescence microscopy. ( Left ) Alexa488-azide stain. ( Center ) Alexa594-azide stain. ( Right ) Overlay of the Alexa488 and Alexa594 images. The complete colocalization of the two colors indicates that the two successive azide stains detect ethynyl groups that have the same distribution within the cell nucleus. ( B ) NIH 3T3 cells labeled by incubation overnight with 2 μM EdU and 2 μM BrdU were fixed and reacted with 10 μM Alexa594-azide, followed by standard BrdU immunodetection by using an anti-BrdU monoclonal antibody and an Alexa488-conjugated secondary antibody. ( Left ) BrdU stain. ( Center ) EdU stain. ( Right ) Overlay of the BrdU and EdU images. Note that each cell that incorporated BrdU also incorporated EdU. The BrdU micrograph was taken by using a five-times longer exposure than the exposure used for EdU (500 ms compared with 100 ms) under identical digital camera settings.
    Figure Legend Snippet: Reproducibility of EdU labeling ( A ) and comparison with BrdU ( B ). ( A ) NIH 3T3 cells labeled by incubation overnight with 2 μM EdU were fixed and reacted successively with 10 μM Alexa488-azide and 10 μM Alexa594-azide, respectively, as described in Materials and Methods . The cells were imaged by fluorescence microscopy. ( Left ) Alexa488-azide stain. ( Center ) Alexa594-azide stain. ( Right ) Overlay of the Alexa488 and Alexa594 images. The complete colocalization of the two colors indicates that the two successive azide stains detect ethynyl groups that have the same distribution within the cell nucleus. ( B ) NIH 3T3 cells labeled by incubation overnight with 2 μM EdU and 2 μM BrdU were fixed and reacted with 10 μM Alexa594-azide, followed by standard BrdU immunodetection by using an anti-BrdU monoclonal antibody and an Alexa488-conjugated secondary antibody. ( Left ) BrdU stain. ( Center ) EdU stain. ( Right ) Overlay of the BrdU and EdU images. Note that each cell that incorporated BrdU also incorporated EdU. The BrdU micrograph was taken by using a five-times longer exposure than the exposure used for EdU (500 ms compared with 100 ms) under identical digital camera settings.

    Techniques Used: Labeling, Incubation, Fluorescence, Microscopy, Staining, Immunodetection, Mass Spectrometry

    2) Product Images from "Exdpf Is a Key Regulator of Exocrine Pancreas Development Controlled by Retinoic Acid and ptf1a in Zebrafish"

    Article Title: Exdpf Is a Key Regulator of Exocrine Pancreas Development Controlled by Retinoic Acid and ptf1a in Zebrafish

    Journal: PLoS Biology

    doi: 10.1371/journal.pbio.0060293

    Reduced exdpf Causes Defects of Exocrine Cell Proliferation (A) BrdU labeling result in 33 hpf embryos. Green: MP760 :GFP , Red: BrdU staining. Blue: DAPI staining. Dorsal view, anterior to the left. Enlargement represents higher magnification of boxed area in each row. Arrows indicate non-proliferating cells; arrowheads indicate proliferating cells as evidenced by overlapping of red and green colors. (B) BrdU labeling result in 3 dpf embryos. Green: elastase A:GFP , Red: BrdU staining. Lateral view, anterior to the left. Scale bar: 50 μm. Arrows indicate non-proliferating cells; arrowheads indicate proliferating cells as evidenced by overlapping of red and green colors. (C) Quantitative graphs for BrdU incorporation rate in 33 hpf or 3 dpf embryos. The average number of GFP positive cells with BrdU incorporation was obtained by counting BrdU-labeled GFP positive cells from five embryos. Y axis: Mean ± SD. (D) Semiquantitative RT-PCR examination of expression of cyclin D1 and cell cycle inhibitors p21 , p27 and cyclin G1 in control (control), exdpf morphants (MO), and exdpf mRNA injected embryos (RNA) at 33 hpf, 2 dpf, 3 dpf, and 5 dpf. In each group, 30 embryos were used to extract total RNA for RT-PCR.
    Figure Legend Snippet: Reduced exdpf Causes Defects of Exocrine Cell Proliferation (A) BrdU labeling result in 33 hpf embryos. Green: MP760 :GFP , Red: BrdU staining. Blue: DAPI staining. Dorsal view, anterior to the left. Enlargement represents higher magnification of boxed area in each row. Arrows indicate non-proliferating cells; arrowheads indicate proliferating cells as evidenced by overlapping of red and green colors. (B) BrdU labeling result in 3 dpf embryos. Green: elastase A:GFP , Red: BrdU staining. Lateral view, anterior to the left. Scale bar: 50 μm. Arrows indicate non-proliferating cells; arrowheads indicate proliferating cells as evidenced by overlapping of red and green colors. (C) Quantitative graphs for BrdU incorporation rate in 33 hpf or 3 dpf embryos. The average number of GFP positive cells with BrdU incorporation was obtained by counting BrdU-labeled GFP positive cells from five embryos. Y axis: Mean ± SD. (D) Semiquantitative RT-PCR examination of expression of cyclin D1 and cell cycle inhibitors p21 , p27 and cyclin G1 in control (control), exdpf morphants (MO), and exdpf mRNA injected embryos (RNA) at 33 hpf, 2 dpf, 3 dpf, and 5 dpf. In each group, 30 embryos were used to extract total RNA for RT-PCR.

    Techniques Used: Labeling, BrdU Staining, Staining, BrdU Incorporation Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection

    3) Product Images from "Electrical Stimulation Influences Satellite Cell Proliferation and Apoptosis in Unloading-Induced Muscle Atrophy in Mice"

    Article Title: Electrical Stimulation Influences Satellite Cell Proliferation and Apoptosis in Unloading-Induced Muscle Atrophy in Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030348

    Satellite cell proliferation. (A) Immunohistochemically stained cross-sections of soleus muscle from weight-bearing (WB), hindlimb-unloaded (HU), and electrically stimulated (HU-ES) groups showing expression of Pax7 (red) and BrdU (green) incorporation counterstained with DAPI (blue) for nuclei identification. Yellow staining represents Pax7 and BrdU double-positive nuclei. Scale bar = 25 µm. (B) Co-immunostaining of Pax7 and dystrophin on muscle cross-section for satellite cell localization. A Pax7 + nucleus (green, arrow ) located outside the dystrophin + (red) plasma membrane of a myofibre. DAPI (blue) shows the nuclei. Scale bar = 20 µm. (C) Total Pax7 immunoreactive nuclei; BrdU + /Pax7 + double-positive nuclei per 10 3 myofibers; and ratio of BrdU + /Pax7 + nuclei within the total Pax7 + population. *p
    Figure Legend Snippet: Satellite cell proliferation. (A) Immunohistochemically stained cross-sections of soleus muscle from weight-bearing (WB), hindlimb-unloaded (HU), and electrically stimulated (HU-ES) groups showing expression of Pax7 (red) and BrdU (green) incorporation counterstained with DAPI (blue) for nuclei identification. Yellow staining represents Pax7 and BrdU double-positive nuclei. Scale bar = 25 µm. (B) Co-immunostaining of Pax7 and dystrophin on muscle cross-section for satellite cell localization. A Pax7 + nucleus (green, arrow ) located outside the dystrophin + (red) plasma membrane of a myofibre. DAPI (blue) shows the nuclei. Scale bar = 20 µm. (C) Total Pax7 immunoreactive nuclei; BrdU + /Pax7 + double-positive nuclei per 10 3 myofibers; and ratio of BrdU + /Pax7 + nuclei within the total Pax7 + population. *p

    Techniques Used: Staining, Western Blot, Expressing, Immunostaining

    4) Product Images from "Regeneration of Pancreatic Non-? Endocrine Cells in Adult Mice following a Single Diabetes-Inducing Dose of Streptozotocin"

    Article Title: Regeneration of Pancreatic Non-? Endocrine Cells in Adult Mice following a Single Diabetes-Inducing Dose of Streptozotocin

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0036675

    BrdU incorporation demonstrating that cell proliferation was involved in non-β-cell regeneration. Following STZ injection, 1 mg/ml of BrdU was added into the mice's drinking water. Normal mice were included as the control. At different day post-STZ, the pancreases were processed for immunofluorescence staining with anti-BrdU (red, A and C) and anti-glucagon (green, A) or anti-somatostatin (green, C). The arrows mark BrdU + α-cells (A) and BrdU + δ-cells (C) in the representative islets. The percentage of BrdU + α-cells (B) and BrdU + δ-cells (D) were calculated by dividing the number of these cells by total α- and δ-cells in each islet, respectively. Unpaired t-tests were performed to compare whether the percentage at each time point following STZ treatment was significantly different from that in normal mice. *: p
    Figure Legend Snippet: BrdU incorporation demonstrating that cell proliferation was involved in non-β-cell regeneration. Following STZ injection, 1 mg/ml of BrdU was added into the mice's drinking water. Normal mice were included as the control. At different day post-STZ, the pancreases were processed for immunofluorescence staining with anti-BrdU (red, A and C) and anti-glucagon (green, A) or anti-somatostatin (green, C). The arrows mark BrdU + α-cells (A) and BrdU + δ-cells (C) in the representative islets. The percentage of BrdU + α-cells (B) and BrdU + δ-cells (D) were calculated by dividing the number of these cells by total α- and δ-cells in each islet, respectively. Unpaired t-tests were performed to compare whether the percentage at each time point following STZ treatment was significantly different from that in normal mice. *: p

    Techniques Used: BrdU Incorporation Assay, Injection, Mouse Assay, Immunofluorescence, Staining

    Related Articles

    Incubation:

    Article Title: Exdpf Is a Key Regulator of Exocrine Pancreas Development Controlled by Retinoic Acid and ptf1a in Zebrafish
    Article Snippet: Embryos were placed in 10 mM solution of BrdU (Sigma) in fish water at 24 hpf and kept in dark at 28 °C for 48 h. Then embryos were fixed in 4% PFA for 2 h at room temperature. .. After washing (5 × 5 min in PBST), embryos were incubated in 1 N HCl for 1 h. Then embryos were washed again for 5 × 5 min in PBST and blocked using 5% goat serum for 1 h at room temperature and incubated over night at 4 °C in 1:100 mouse anti-BrdU monoclonal antibody (Chemicon) and 1:500 rabbit anti-GFP polyclonal antibody (Abcam). .. After washing (5 × 5 min in PBST), embryos were incubated overnight at 4 °C in 1:200 A488 conjugated goat anti-rabbit antibody (Molecular Probe).

    Article Title: Ultrastructural Characteristics of 5BrdU Labeling Retention Cells Including Stem Cells of Regenerating Feathers in Chicken
    Article Snippet: .. The sections were pretreated with 0.2 mol l−1 HCl for 2 h, rinsed in buffer, and incubated overnight at 0–4°C with monoclonal anti-5BrdU (Sigma or Chemicon), diluted 1:100 in 0.05 Tris buffer with 1% bovine serum albumine, at pH 7.2. .. Sections were rinsed in the buffer and incubated for 1 h at room temperature with secondary Biotinylated Goat anti Mouse IgG antibody (Vector Laboratories, 1:200 dilution) for 1 h and third antibody Streptavidin (Vector Laboratories, 1:200 dilution) for 30 min. AEC substrate kit (Vector Laboratories) was used to develop staining.

    Article Title: Electrical Stimulation Influences Satellite Cell Proliferation and Apoptosis in Unloading-Induced Muscle Atrophy in Mice
    Article Snippet: Fixed sections were incubated with 2N HCl for DNA denaturation. .. After several washes, the sections were incubated with mouse anti-BrdU monoclonal antibody (1∶100; Sigma-Aldrich, USA) and rabbit anti-Pax7 polyclonal antibody (1∶100; catalog no. ab34360, Abcam, USA) at 4°C overnight, followed by Alexa 488-conjugated donkey anti-mouse IgG and Alexa 556-conjugated donkey anti-rabbit IgG (1∶200; Invitrogen). .. To confirm successful BrdU incorporation, the skin tissue was harvested and sectioned as a positive control.

    Article Title: Neuroimmune and epigenetic involvement in adolescent binge ethanol-induced loss of basal forebrain cholinergic neurons: Restoration with voluntary exercise
    Article Snippet: .. Briefly, sections were incubated in a primary antibody solution containing goat anti‐ChAT (Millipore, Temecula, CA, Cat. #AB144P), rabbit anti‐TrkA (Millipore, Cat. #06‐574), mouse anti‐p75NTR (Millipore, Cat. #MAB365), mouse anti‐NeuN (Millipore, Cat. #MAB377), rabbit antiphosphorylated NF‐κB p65 Ser 536 (pNF‐κB p65; Abcam, Cambridge, MA, Cat. #ab86299), or mouse anti‐BrdU (Millipore, Cat. #MAB3424) for 24 hours at 4°C. .. For BrdU immunohistochemistry, DNA was additionally denatured by incubation in 2 N HCl for 30 minutes at 37°C followed by incubation in 0.1 M boric acid for 10 minutes at room temperature (pH: 8.5).

    other:

    Article Title: Regeneration of Pancreatic Non-? Endocrine Cells in Adult Mice following a Single Diabetes-Inducing Dose of Streptozotocin
    Article Snippet: Antibodies Guinea pig anti-human Insulin polyclonal antibody (1∶100), mouse anti-BrdU monoclonal antibody (1∶500), rabbit anti-Glucagon (1∶200), rabbit anti-Somatostatin (1∶200), and rabbit anti-PPP (1∶300) polyclonal antibodies were purchased from Millipore Corp. (Billerica, Massachusetts).

    Immunoprecipitation:

    Article Title: New mitochondrial DNA synthesis enables NLRP3 inflammasome activation
    Article Snippet: .. For detection of BrdU and 8-OH-dG in the ASC immunoprecipitation products, the eluted samples were dot-blotted and UV cross-linked to a nitrocellulose membrane that was immunoblotted with BrdU monoclonal antibody (BU33; Sigma) or 8OH-dG BrdU monoclonal antibody (15A3; Rockland Immunochemicals). .. For the detection of NLRP3, AIM2, ASC and pro-caspase-1 in the ASC immunoprecipitation products, the eluted samples were heated to 95 °C for 5 min, the gel was separated by SDS-PAGE, transferred to a nitrocellulose membrane and immunoblotted with antibodies against ASC (2EI-7, Millipore), NLRP3 (AG-20B-0014-C100, Adipogen), pro-casp1 (AG-20B-0042-C100, Adipogen) and AIM2 (sc-137967, Santa Cruz Biotechnology).

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    Millipore antihuman bmi 1 monoclonal antibody
    Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 <t>(BMI-1)</t> antigen in human retinoblastoma retinae. (Upper panel) Undifferentiated retinoblastomas and (lower panel) differentiated retinoblastomas grouped according to TNM classification. Some specimens had an indistinct inner nuclear layer (marked by red brackets). There was no consistent difference of BMI-1 expression in retinas of undifferentiated and differentiated retinoblastomas. Scale bars: 50 μm.
    Antihuman Bmi 1 Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antihuman bmi 1 monoclonal antibody/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antihuman bmi 1 monoclonal antibody - by Bioz Stars, 2021-04
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    97
    Millipore mouse anti brdu
    Combination therapy improved the transplantation and differentiation of BMSCs in ischemic brain. (a) Immunofluorescence staining for <t>BrdU</t> and <t>Nestin</t> positive cells at day 7 after MCAO. BrdU (red, 1) and Nestin (green, 2) in BMSCs monotherapy group, BrdU (red, 4) and Nestin (green, 5) in the combination group. Panels 3 and 6 are the merged images, respectively. Arrows represent the double-immunofluorescence cells. Scale bar indicates 200 μm in BMSC and combination group. (b) The number of engrafted-BMSCs (i) and Nestin positive cells (ii) in BMSCs monotherapy group and combination group. * P
    Mouse Anti Brdu, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti brdu/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    97
    Millipore monoclonal antibody recognizing γ tubulin
    Centrosome Duplication Is Uncoupled from DNA Replication in vHMECs (A) Schematic of the centrosome duplication cycle (interior, green) in relation to the DNA replication cycle (exterior, blue). Each daughter cell inherits one copy of the DNA and one centrosome (G1 phase of the cell cycle). Centrosomes (small, green circles) begin duplication at the same point as when DNA replication is initiated (S phase of the cell cycle). Following duplication and maturation of the centrosome, the two centrosomes separate and migrate to opposite poles during early M (mitosis). (B) Analysis of mononucleated cells with more than two centrosomes in HMECs (black: RM9 [5 PD], RM16 [less than 4 PD]), early-passage vHMECs (red: RM9 [21 PD], RM16 [7 PD]), and low FSC/SSC sorted vHMECs (blue: RM16 [17 and 30 PD]) untreated (−HU) or exposed to HU (+HU). (C–E) Examples of normal centrosome numbers in HMECs (C) and vHMECs (D) and more than two centrosomes in vHMECs (E). (F) Early-passage vHMECs (RM9 [14 PD] and RM16 [17 PD]) that were exposed to HU were stained with an antibody recognizing <t>γ-tubulin</t> and with PI (DNA counterstain), and the DNA content of each nucleus was measured by quantitative immunofluorescence microscopy. Cells were classified as having 2N to 4N (diploid) or more than 4N (polyploidy) DNA content. The centrosome number of each cell was correlated to the DNA content of that cell. Analysis included 100 to 200 cells (excluding binucleated cells). *Statistical significance ( p
    Monoclonal Antibody Recognizing γ Tubulin, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 97 stars, based on 1 article reviews
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    97
    Millipore rat monoclonal anti brdu
    Olfactory discrimination learning preferentially recruits 5-week-old neurons in the OB. A , Confocal illustration showing <t>BrdU-IR</t> cells (red) coexpressing the neuronal marker NeuN (blue) and the cellular activation factor <t>Fos</t> (green). B , Percentage of adult-born neurons expressing Fos in the GCL. C , Number of Fos-IR cells, n = 5–8 per group, and illustration of Fos-IR cells in the GCL on the right top. D , Percentage of preexisting and adult-born (same data as in B ) neurons expressing Fos in the GCL. Both means and individual data are presented. ** p
    Rat Monoclonal Anti Brdu, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat monoclonal anti brdu/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in human retinoblastoma retinae. (Upper panel) Undifferentiated retinoblastomas and (lower panel) differentiated retinoblastomas grouped according to TNM classification. Some specimens had an indistinct inner nuclear layer (marked by red brackets). There was no consistent difference of BMI-1 expression in retinas of undifferentiated and differentiated retinoblastomas. Scale bars: 50 μm.

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in human retinoblastoma retinae. (Upper panel) Undifferentiated retinoblastomas and (lower panel) differentiated retinoblastomas grouped according to TNM classification. Some specimens had an indistinct inner nuclear layer (marked by red brackets). There was no consistent difference of BMI-1 expression in retinas of undifferentiated and differentiated retinoblastomas. Scale bars: 50 μm.

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Light Microscopy, Staining, Expressing

    Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in differentiated retinoblastomas grouped according to Tumor, Nodes, Metastasis (TNM) classification. A : Hematoxylin and eosin staining. Closed red arrows in A show the Flexner-Wintersteiner rosettes. B: Magnified images in C : weak to moderate BMI-1 expression in differentiated retinoblastomas. D : Background staining of BMI-1 in retinoblastoma 8927 with incubation of primary antibody. E : Negligible BMI-1 expression in a normal human retina section. Scale bars: 50 µm.

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in differentiated retinoblastomas grouped according to Tumor, Nodes, Metastasis (TNM) classification. A : Hematoxylin and eosin staining. Closed red arrows in A show the Flexner-Wintersteiner rosettes. B: Magnified images in C : weak to moderate BMI-1 expression in differentiated retinoblastomas. D : Background staining of BMI-1 in retinoblastoma 8927 with incubation of primary antibody. E : Negligible BMI-1 expression in a normal human retina section. Scale bars: 50 µm.

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Light Microscopy, Staining, Expressing, Incubation

    Influence of B lymphoma Mo-MLV insertion region 1 (BMI-1) on apoptosis and gene expression of Y79 cells. A : TUNEL assay showing increase of apoptosis rate after siBmi1 transfection to Y79 cells. A mild reduction of apoptosis was seen with BMI-1-HA expression (n=3, mean±standard deviation). *p

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: Influence of B lymphoma Mo-MLV insertion region 1 (BMI-1) on apoptosis and gene expression of Y79 cells. A : TUNEL assay showing increase of apoptosis rate after siBmi1 transfection to Y79 cells. A mild reduction of apoptosis was seen with BMI-1-HA expression (n=3, mean±standard deviation). *p

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Expressing, TUNEL Assay, Transfection, Standard Deviation

    B lymphoma Mo-MLV insertion region 1 (BMI-1) expression altered retinoblastoma 480 Y79 cell growth. A - C : Immunostaining and reverse transcription (RT)–PCR analyses show marked increase of BMI-1 transcript and protein in Y79 cells after transfection with pCMV-HA/BMI-1 compared with vector control. D and E : MTT assay showing Y79 cell viability and proliferation was promoted by BMI-1 overexpression and reduced by siRNA-mediated inhibition of Bmi-1 . F : Gene expression analysis of p14, p16, and PCNA in pCMV-HA/BMI-1, pSuper-BMI-1-si, and vector-transfected Y79 cells. *p

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: B lymphoma Mo-MLV insertion region 1 (BMI-1) expression altered retinoblastoma 480 Y79 cell growth. A - C : Immunostaining and reverse transcription (RT)–PCR analyses show marked increase of BMI-1 transcript and protein in Y79 cells after transfection with pCMV-HA/BMI-1 compared with vector control. D and E : MTT assay showing Y79 cell viability and proliferation was promoted by BMI-1 overexpression and reduced by siRNA-mediated inhibition of Bmi-1 . F : Gene expression analysis of p14, p16, and PCNA in pCMV-HA/BMI-1, pSuper-BMI-1-si, and vector-transfected Y79 cells. *p

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Expressing, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, MTT Assay, Over Expression, Inhibition

    Effect of B lymphoma Mo-MLV insertion 486 region 1 (BMI-1) on multicellular sphere formation of Y79 cells. A : Single transfected cells were cultured for 7 days to form multicellular spheres. The number and size of spheres were quantified. In triplicated experiments, the mean percentage of spheres having with diameters was indicated. B and C : Cell proliferation assay by BrdU incorporation for Y79 cells transfected with BMI-1-HA. B : BMI-1-HA and C : siBmi 1. Scale bar: 100 μm.

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: Effect of B lymphoma Mo-MLV insertion 486 region 1 (BMI-1) on multicellular sphere formation of Y79 cells. A : Single transfected cells were cultured for 7 days to form multicellular spheres. The number and size of spheres were quantified. In triplicated experiments, the mean percentage of spheres having with diameters was indicated. B and C : Cell proliferation assay by BrdU incorporation for Y79 cells transfected with BMI-1-HA. B : BMI-1-HA and C : siBmi 1. Scale bar: 100 μm.

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Transfection, Cell Culture, Proliferation Assay, BrdU Incorporation Assay

    Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in undifferentiated retinoblastomas grouped according to Tumor, Nodes, Metastasis (TNM) classification. A : Retinoblastoma sections were stained by hematoxylin and eosin. B: Magnified images in C : intense nuclear BMI-1 expression in undifferentiated retinoblastomas. Scale bars: 50 µm.

    Journal: Molecular Vision

    Article Title: Role of B lymphoma Mo-MLV insertion region 1 in the oncogenic behavior of retinoblastomas

    doi:

    Figure Lengend Snippet: Representative light microscopy pictures of staining of B lymphoma Mo-MLV insertion region 1 (BMI-1) antigen in undifferentiated retinoblastomas grouped according to Tumor, Nodes, Metastasis (TNM) classification. A : Retinoblastoma sections were stained by hematoxylin and eosin. B: Magnified images in C : intense nuclear BMI-1 expression in undifferentiated retinoblastomas. Scale bars: 50 µm.

    Article Snippet: After blocking with 5% normal goat serum and 0.1% Triton X-100, the section was incubated with or without 0.5 to 1 μg/ml antihuman BMI-1 monoclonal antibody (Clone F6, Millipore, Billerica, MA).

    Techniques: Light Microscopy, Staining, Expressing

    Combination therapy improved the transplantation and differentiation of BMSCs in ischemic brain. (a) Immunofluorescence staining for BrdU and Nestin positive cells at day 7 after MCAO. BrdU (red, 1) and Nestin (green, 2) in BMSCs monotherapy group, BrdU (red, 4) and Nestin (green, 5) in the combination group. Panels 3 and 6 are the merged images, respectively. Arrows represent the double-immunofluorescence cells. Scale bar indicates 200 μm in BMSC and combination group. (b) The number of engrafted-BMSCs (i) and Nestin positive cells (ii) in BMSCs monotherapy group and combination group. * P

    Journal: Chinese Medical Journal

    Article Title: Possible Mechanism of Therapeutic Effect of 3-Methyl-1-phenyl-2-pyrazolin-5-one and Bone Marrow Stromal Cells Combination Treatment in Rat Ischemic Stroke Model

    doi: 10.4103/0366-6999.183418

    Figure Lengend Snippet: Combination therapy improved the transplantation and differentiation of BMSCs in ischemic brain. (a) Immunofluorescence staining for BrdU and Nestin positive cells at day 7 after MCAO. BrdU (red, 1) and Nestin (green, 2) in BMSCs monotherapy group, BrdU (red, 4) and Nestin (green, 5) in the combination group. Panels 3 and 6 are the merged images, respectively. Arrows represent the double-immunofluorescence cells. Scale bar indicates 200 μm in BMSC and combination group. (b) The number of engrafted-BMSCs (i) and Nestin positive cells (ii) in BMSCs monotherapy group and combination group. * P

    Article Snippet: Sections were incubated in 2 mol/L HCl at 37°C for 30 min to denature DNA and exposed BrdU, rinsed thoroughly in PBS and incubated in a mixture of mouse anti-BrdU (1:100, Sigma, USA) and rabbit anti-Nestin (1:100, Sigma, USA) at room temperature for 2 h. After incubation, sections were washed in PBS and placed for 2 h in tetramethyl rhodamine isothiocyanate-conjugated goat anti-mouse IgG (1:100, Sigma, USA) for BrdU and FITC-conjugated goat anti-rabbit IgG (1:100, Sigma, USA) for Nestin.

    Techniques: Transplantation Assay, Immunofluorescence, Staining

    Centrosome Duplication Is Uncoupled from DNA Replication in vHMECs (A) Schematic of the centrosome duplication cycle (interior, green) in relation to the DNA replication cycle (exterior, blue). Each daughter cell inherits one copy of the DNA and one centrosome (G1 phase of the cell cycle). Centrosomes (small, green circles) begin duplication at the same point as when DNA replication is initiated (S phase of the cell cycle). Following duplication and maturation of the centrosome, the two centrosomes separate and migrate to opposite poles during early M (mitosis). (B) Analysis of mononucleated cells with more than two centrosomes in HMECs (black: RM9 [5 PD], RM16 [less than 4 PD]), early-passage vHMECs (red: RM9 [21 PD], RM16 [7 PD]), and low FSC/SSC sorted vHMECs (blue: RM16 [17 and 30 PD]) untreated (−HU) or exposed to HU (+HU). (C–E) Examples of normal centrosome numbers in HMECs (C) and vHMECs (D) and more than two centrosomes in vHMECs (E). (F) Early-passage vHMECs (RM9 [14 PD] and RM16 [17 PD]) that were exposed to HU were stained with an antibody recognizing γ-tubulin and with PI (DNA counterstain), and the DNA content of each nucleus was measured by quantitative immunofluorescence microscopy. Cells were classified as having 2N to 4N (diploid) or more than 4N (polyploidy) DNA content. The centrosome number of each cell was correlated to the DNA content of that cell. Analysis included 100 to 200 cells (excluding binucleated cells). *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Centrosome Duplication Is Uncoupled from DNA Replication in vHMECs (A) Schematic of the centrosome duplication cycle (interior, green) in relation to the DNA replication cycle (exterior, blue). Each daughter cell inherits one copy of the DNA and one centrosome (G1 phase of the cell cycle). Centrosomes (small, green circles) begin duplication at the same point as when DNA replication is initiated (S phase of the cell cycle). Following duplication and maturation of the centrosome, the two centrosomes separate and migrate to opposite poles during early M (mitosis). (B) Analysis of mononucleated cells with more than two centrosomes in HMECs (black: RM9 [5 PD], RM16 [less than 4 PD]), early-passage vHMECs (red: RM9 [21 PD], RM16 [7 PD]), and low FSC/SSC sorted vHMECs (blue: RM16 [17 and 30 PD]) untreated (−HU) or exposed to HU (+HU). (C–E) Examples of normal centrosome numbers in HMECs (C) and vHMECs (D) and more than two centrosomes in vHMECs (E). (F) Early-passage vHMECs (RM9 [14 PD] and RM16 [17 PD]) that were exposed to HU were stained with an antibody recognizing γ-tubulin and with PI (DNA counterstain), and the DNA content of each nucleus was measured by quantitative immunofluorescence microscopy. Cells were classified as having 2N to 4N (diploid) or more than 4N (polyploidy) DNA content. The centrosome number of each cell was correlated to the DNA content of that cell. Analysis included 100 to 200 cells (excluding binucleated cells). *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Staining, Immunofluorescence, Microscopy

    Loss of p16 INK4a Results in Centrosome Dysfunction and the Subsequent Generation of Aneuploid HMF and HeLa Cells (A) Western blot analysis of p16 INK4a expression in HMF and HeLa cells infected with vector-only (vector) or shRNA directed against p16 INK4a (p16 INK4a shRNA). (B) Examples of more than two centrosomes in HMF (p16 INK4a shRNA) and HeLa (p16 INK4a shRNA) cells. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein. (C) Analysis of parental HMF (RM9 [1 PD], RM21 [3 PD]) and HeLa cells (black) and HMF and HeLa cells infected with vector-only (HMF: RM9 [7 PD], RM21 [11 PD]) (white) or HMF and HeLa cells infected with p16 INK4a shRNA (HMF: RM9 [6 PD], RM21 [7 PD]) (gray) containing mononucleated cells with more than two centrosomes. Cells were untreated (−HU) or exposed to HU (+HU). Analysis included more than two HMF and HeLa cells. *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Loss of p16 INK4a Results in Centrosome Dysfunction and the Subsequent Generation of Aneuploid HMF and HeLa Cells (A) Western blot analysis of p16 INK4a expression in HMF and HeLa cells infected with vector-only (vector) or shRNA directed against p16 INK4a (p16 INK4a shRNA). (B) Examples of more than two centrosomes in HMF (p16 INK4a shRNA) and HeLa (p16 INK4a shRNA) cells. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein. (C) Analysis of parental HMF (RM9 [1 PD], RM21 [3 PD]) and HeLa cells (black) and HMF and HeLa cells infected with vector-only (HMF: RM9 [7 PD], RM21 [11 PD]) (white) or HMF and HeLa cells infected with p16 INK4a shRNA (HMF: RM9 [6 PD], RM21 [7 PD]) (gray) containing mononucleated cells with more than two centrosomes. Cells were untreated (−HU) or exposed to HU (+HU). Analysis included more than two HMF and HeLa cells. *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Western Blot, Expressing, Infection, Plasmid Preparation, shRNA, Immunocytochemistry

    Supernumerary Centrosomes Produce Multipolar Spindles and Correlate with Aneuploidy (A) BrdU incorporation of HMECs (top) and vHMECs (bottom). Cell cycle analysis were performed using BrdU incorporation for cells that were untreated (−HU), treated for 48 h with HU (+HU), or treated for 48 h with HU, followed by release from HU treatment for 7 to 8 d (+HU → release). DNA replication resumes following release from 48 h of HU treatment. (B) Microtubule nucleation sites were determined by immunocytochemistry with an antibody recognizing γ-tubulin. (C) Analysis of HMECs (black: RM9 and RM26 [less than 4 PD]) and vHMECs (red: RM15 [25 PD], RM16 [13 PD]) for multipolar mitosis in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release). (D) Analysis of HMECs (black: RM9 and RM15 [less than 5 PD]) and vHMECs (red: RM9 [21 PD], RM16 [17 PD]), HMECs infected with p16 INK4a shRNA (gray) for genomic abnormalities in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release). Types of chromosomal abnormalities represented include aneuploidy, structural abnormalities, and telomeric associations. Analysis included at least 100 metaphases. *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Supernumerary Centrosomes Produce Multipolar Spindles and Correlate with Aneuploidy (A) BrdU incorporation of HMECs (top) and vHMECs (bottom). Cell cycle analysis were performed using BrdU incorporation for cells that were untreated (−HU), treated for 48 h with HU (+HU), or treated for 48 h with HU, followed by release from HU treatment for 7 to 8 d (+HU → release). DNA replication resumes following release from 48 h of HU treatment. (B) Microtubule nucleation sites were determined by immunocytochemistry with an antibody recognizing γ-tubulin. (C) Analysis of HMECs (black: RM9 and RM26 [less than 4 PD]) and vHMECs (red: RM15 [25 PD], RM16 [13 PD]) for multipolar mitosis in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release). (D) Analysis of HMECs (black: RM9 and RM15 [less than 5 PD]) and vHMECs (red: RM9 [21 PD], RM16 [17 PD]), HMECs infected with p16 INK4a shRNA (gray) for genomic abnormalities in untreated (−HU) or exposed to HU followed by release from HU treatment (+HU → release). Types of chromosomal abnormalities represented include aneuploidy, structural abnormalities, and telomeric associations. Analysis included at least 100 metaphases. *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: BrdU Incorporation Assay, Cell Cycle Assay, Immunocytochemistry, Infection, shRNA

    Unregulated Kinase Activity Is Responsible for the Uncoupling of the Centrosome Duplication and DNA Replication Cycles (A and B) vHMECs (RM9 [17 PD], RM16 [20 PD]) that were untreated (−HU) or treated for 6, 12, 24, 36, and 48 h with HU (+HU) were analyzed by (A) flow cytometry of PI and BrdU incorporation (% cells arrested in S phase) and (B) confocal microscopy for mononucleated cells with more than two centrosomes. The kinase inhibitors purvalanol A (Pur) and roscovitine (Rosc) were added following the addition of HU for 36 h and incubated in the presence of HU for 12 h (B). (C) Fibroblasts that express p21 (p21 +/+ ) and that have p21 knocked out (p21 −/− ) were analyzed by confocal microscopy for mononucleated cells with more than two centrosomes. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome associated γ-tubulin protein. Centrosome number analysis included at least 200 cells per sample (excluding binucleated cells). *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Unregulated Kinase Activity Is Responsible for the Uncoupling of the Centrosome Duplication and DNA Replication Cycles (A and B) vHMECs (RM9 [17 PD], RM16 [20 PD]) that were untreated (−HU) or treated for 6, 12, 24, 36, and 48 h with HU (+HU) were analyzed by (A) flow cytometry of PI and BrdU incorporation (% cells arrested in S phase) and (B) confocal microscopy for mononucleated cells with more than two centrosomes. The kinase inhibitors purvalanol A (Pur) and roscovitine (Rosc) were added following the addition of HU for 36 h and incubated in the presence of HU for 12 h (B). (C) Fibroblasts that express p21 (p21 +/+ ) and that have p21 knocked out (p21 −/− ) were analyzed by confocal microscopy for mononucleated cells with more than two centrosomes. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome associated γ-tubulin protein. Centrosome number analysis included at least 200 cells per sample (excluding binucleated cells). *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Activity Assay, Flow Cytometry, Cytometry, BrdU Incorporation Assay, Confocal Microscopy, Incubation, Immunocytochemistry

    vHMECs Accumulate Mitotic and Centrosome Abnormalities (A) The solid line graph represents the in vitro growth curves of both HMECs (black circles) and vHMECs (red squares) isolated from RM16. The bar graph represents analysis of mononucleated cells containing more than two centrosomes. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein (excluding multinucleated cells). Cells were analyzed at multiple points along the growth curves of two different individuals (RM9 and RM16). HMEC (black) (RM9 and RM16 [less than five PD]) and vHMECs (red) analyzed at early-passage (RM9 [14 PD] and RM16 [20 PD]), late-passage (RM9 [43 PD], RM9 [65 PD]), and the agonescence (RM9 [70 PD] and RM16 [50 PD]). *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: vHMECs Accumulate Mitotic and Centrosome Abnormalities (A) The solid line graph represents the in vitro growth curves of both HMECs (black circles) and vHMECs (red squares) isolated from RM16. The bar graph represents analysis of mononucleated cells containing more than two centrosomes. Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein (excluding multinucleated cells). Cells were analyzed at multiple points along the growth curves of two different individuals (RM9 and RM16). HMEC (black) (RM9 and RM16 [less than five PD]) and vHMECs (red) analyzed at early-passage (RM9 [14 PD] and RM16 [20 PD]), late-passage (RM9 [43 PD], RM9 [65 PD]), and the agonescence (RM9 [70 PD] and RM16 [50 PD]). *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: In Vitro, Isolation, Immunocytochemistry

    Supernumerary Centrosomes Play a Causal Role in the Production of Aneuploid Cells (A) Early-passage vHMECs (RM18, 13–33 PD) that express EGFP–γ-tubulin (green) and EGFP-H2B (green) were stained with antibody recognizing the centrosome associated γ-tubulin protein (red) and with DAPI to localize DNA (blue). The merged image demonstrates colocalization of EGFP–γ-tubulin with the centrosome and EGFP-H2B with the DNA. (B) Examples of the mitotic progression of cells that divide into two nuclei with two centrosomes (−HU and +HU → release; bipolar, two centrosomes), that divide into two nuclei with more than two centrosomes (+HU → release; bipolar, more than two centrosomes), and that divide into greater than two nuclei with more than two centrosomes (+HU → release; multipolar, with more than two centrosomes). Arrowhead points to the EGFP–γ-tubulin signal (centrosomes). The EGFP-H2B signal was selected (pastel-colored nuclei) and the total signal intensity was quantitated. Determining the fold difference (EGFP-H2B signal intensity of daughter cells 1/EGFP-H2B signal intensity of daughter cells 2) allowed us to determine whether cells had segregated their DNA equally (fold difference close to 1.00) or unequally (fold difference > 1.00). (C) Bar graph of the individual (white, black, red, and red stripe) and mean (yellow) fold differences in EGFP-H2B signals intensity between daughter cells. Standard deviations represent analysis of up to ten time frames per mitotic event. The dashed line represents the average mean fold difference (1.08) of the normal mitosis (−HU and +HU → release; bipolar, two centrosomes). *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Supernumerary Centrosomes Play a Causal Role in the Production of Aneuploid Cells (A) Early-passage vHMECs (RM18, 13–33 PD) that express EGFP–γ-tubulin (green) and EGFP-H2B (green) were stained with antibody recognizing the centrosome associated γ-tubulin protein (red) and with DAPI to localize DNA (blue). The merged image demonstrates colocalization of EGFP–γ-tubulin with the centrosome and EGFP-H2B with the DNA. (B) Examples of the mitotic progression of cells that divide into two nuclei with two centrosomes (−HU and +HU → release; bipolar, two centrosomes), that divide into two nuclei with more than two centrosomes (+HU → release; bipolar, more than two centrosomes), and that divide into greater than two nuclei with more than two centrosomes (+HU → release; multipolar, with more than two centrosomes). Arrowhead points to the EGFP–γ-tubulin signal (centrosomes). The EGFP-H2B signal was selected (pastel-colored nuclei) and the total signal intensity was quantitated. Determining the fold difference (EGFP-H2B signal intensity of daughter cells 1/EGFP-H2B signal intensity of daughter cells 2) allowed us to determine whether cells had segregated their DNA equally (fold difference close to 1.00) or unequally (fold difference > 1.00). (C) Bar graph of the individual (white, black, red, and red stripe) and mean (yellow) fold differences in EGFP-H2B signals intensity between daughter cells. Standard deviations represent analysis of up to ten time frames per mitotic event. The dashed line represents the average mean fold difference (1.08) of the normal mitosis (−HU and +HU → release; bipolar, two centrosomes). *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Staining

    Loss of p16 INK4a Uncouples the Centrosome Duplication and DNA Replication Cycles in HMECs (A) Western blot analysis of p16 INK4a expression in HMECs parental, infected with vector-only (vector), or shRNA directed against p16 INK4a (p16 INK4a shRNA). (B, arrowhead) Examples of more than two centrosomes in HU-exposed HMECs (p16 INK4a shRNA). (C) HMECs infected with vector-only (black: RM9 [6 and 7 PD]) or p16 INK4a shRNA (gray: RM9 [7 and 8 PD]) were untreated (−HU) or exposed to HU (+HU). (D) Expression levels of p16 INK4a (red) in HMECs, vHMECs transfected with vector-only or p16 INK4a . (E) vHMECs infected with vector-only (red: RM15 [33 to 37 PD]) or p16 INK4a (gray: RM15 [33 to 37 PD]) were untreated (−HU) or exposed to HU (+HU). Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein. Analysis included at least 100 cells. *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Loss of p16 INK4a Uncouples the Centrosome Duplication and DNA Replication Cycles in HMECs (A) Western blot analysis of p16 INK4a expression in HMECs parental, infected with vector-only (vector), or shRNA directed against p16 INK4a (p16 INK4a shRNA). (B, arrowhead) Examples of more than two centrosomes in HU-exposed HMECs (p16 INK4a shRNA). (C) HMECs infected with vector-only (black: RM9 [6 and 7 PD]) or p16 INK4a shRNA (gray: RM9 [7 and 8 PD]) were untreated (−HU) or exposed to HU (+HU). (D) Expression levels of p16 INK4a (red) in HMECs, vHMECs transfected with vector-only or p16 INK4a . (E) vHMECs infected with vector-only (red: RM15 [33 to 37 PD]) or p16 INK4a (gray: RM15 [33 to 37 PD]) were untreated (−HU) or exposed to HU (+HU). Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein. Analysis included at least 100 cells. *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Western Blot, Expressing, Infection, Plasmid Preparation, shRNA, Transfection, Immunocytochemistry

    Generation of More Than Two Centrosomes in vHMECs following S Phase Arrest Is Due to Centriole Pair Splitting (A and B) Analysis of the centriole number in of the supernumerary centrosomes of early-passage vHMECs (RM15 [19 PD]) that express EGFP-CETN2 (centriole marker, green) untreated (−HU) or exposed to HU (+HU). Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein (centrosome marker, red). Analysis included at least 100 cells (excluding binucleated cells). *Statistical significance ( p

    Journal: PLoS Biology

    Article Title: p16INK4a Prevents Centrosome Dysfunction and Genomic Instability in Primary CellsA Key Tumor Suppressor Protein Ensures Proper Chromosome Segregation during Cell Division

    doi: 10.1371/journal.pbio.0040051

    Figure Lengend Snippet: Generation of More Than Two Centrosomes in vHMECs following S Phase Arrest Is Due to Centriole Pair Splitting (A and B) Analysis of the centriole number in of the supernumerary centrosomes of early-passage vHMECs (RM15 [19 PD]) that express EGFP-CETN2 (centriole marker, green) untreated (−HU) or exposed to HU (+HU). Centrosome number was determined by immunocytochemistry with an antibody recognizing the centrosome-associated γ-tubulin protein (centrosome marker, red). Analysis included at least 100 cells (excluding binucleated cells). *Statistical significance ( p

    Article Snippet: A monoclonal antibody recognizing γ-tubulin (clone DM1A; Sigma) was used at 1:1,000 dilution to immunostain microtubules.

    Techniques: Marker, Immunocytochemistry

    Olfactory discrimination learning preferentially recruits 5-week-old neurons in the OB. A , Confocal illustration showing BrdU-IR cells (red) coexpressing the neuronal marker NeuN (blue) and the cellular activation factor Fos (green). B , Percentage of adult-born neurons expressing Fos in the GCL. C , Number of Fos-IR cells, n = 5–8 per group, and illustration of Fos-IR cells in the GCL on the right top. D , Percentage of preexisting and adult-born (same data as in B ) neurons expressing Fos in the GCL. Both means and individual data are presented. ** p

    Journal: The Journal of Neuroscience

    Article Title: A Critical Time Window for the Recruitment of Bulbar Newborn Neurons by Olfactory Discrimination Learning

    doi: 10.1523/JNEUROSCI.3941-10.2011

    Figure Lengend Snippet: Olfactory discrimination learning preferentially recruits 5-week-old neurons in the OB. A , Confocal illustration showing BrdU-IR cells (red) coexpressing the neuronal marker NeuN (blue) and the cellular activation factor Fos (green). B , Percentage of adult-born neurons expressing Fos in the GCL. C , Number of Fos-IR cells, n = 5–8 per group, and illustration of Fos-IR cells in the GCL on the right top. D , Percentage of preexisting and adult-born (same data as in B ) neurons expressing Fos in the GCL. Both means and individual data are presented. ** p

    Article Snippet: Briefly, for each staining, one in 10 free-floating sections were incubated with a rat monoclonal anti-BrdU (1/1000, Accurate) or a rabbit polyclonal anti-Fos antibody (1/4000, Calbiochem).

    Techniques: Marker, Activation Assay, Expressing

    Long-term memory recall does not recruit 9-week-old newborn granular neurons exposed to learning at 5 weeks of age. A , Experimental design of the long-term memory recall task. B , C , Percentage of correct responses during the discrimination learning phase ( B ) and during memory recall ( C ). D , Number of BrdU-IR cells. E , Percentage of BrdU-Fos-IR cells in the GCL. C, U-Recall, P-No Recall, P-Recall, n = 5–6 per group; * p

    Journal: The Journal of Neuroscience

    Article Title: A Critical Time Window for the Recruitment of Bulbar Newborn Neurons by Olfactory Discrimination Learning

    doi: 10.1523/JNEUROSCI.3941-10.2011

    Figure Lengend Snippet: Long-term memory recall does not recruit 9-week-old newborn granular neurons exposed to learning at 5 weeks of age. A , Experimental design of the long-term memory recall task. B , C , Percentage of correct responses during the discrimination learning phase ( B ) and during memory recall ( C ). D , Number of BrdU-IR cells. E , Percentage of BrdU-Fos-IR cells in the GCL. C, U-Recall, P-No Recall, P-Recall, n = 5–6 per group; * p

    Article Snippet: Briefly, for each staining, one in 10 free-floating sections were incubated with a rat monoclonal anti-BrdU (1/1000, Accurate) or a rabbit polyclonal anti-Fos antibody (1/4000, Calbiochem).

    Techniques: