rat anti-brdu Search Results


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  • 99
    Millipore rat anti brdu
    Adult hippocampus and OB of D2 KO mice deficient in neurogenesis. (a–d) Extensive labeling in hippocampal DG in controls: wild-type (WT) from D1 KO line (c), WT from D2 KO line (d) and D1 KO mouse (a). Lack of labeling in DG of D2 KO hippocampus (b). (e–h) Similar phenomenon in OB. (i and j) Colocalization (arrows) of <t>BrdU</t> (red) and <t>NeuN</t> (green) in WT hippocampus. (k and l) BrdU-positive nuclei in D2 KO hippocampus never colocalize with NeuN. (o) Similar colocalization (arrowheads indicating green rings) of BrdU (red) and Tuj-1 (green) with no colocalization in D2 KO (p). (m and n) Colabeling of BrdU with glial fibrillary acidic protein (GFAP, green) in D2 KO hippocampus. Confocal images: single-plane (i–l and o and p) and composed (m and n). Bars: (a–h) 200 μm; (i–l, m and n, and o and p) 20 μm.
    Rat Anti Brdu, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 557 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rat anti brdu
    Aged satellite cells cycle more frequently during homeostasis a , Doxycycline (Dox) loading <t>and</t> chase strategy of TetO-H2B-GFP mice and FACS plots for No chase and 84-week chase. SC pool has LRC and nonLRC subsets. b , Strategy and FACS plots for aged (Ag) or adult (Ad) no chase and 12-week chase SCs. Vehicle treated SCs (Neg) (n=10,000 cells,6 animals/group). c , <t>BrdU</t> feeding and representative images of SCs stained with anti-Pax7, BrdU and Dapi. d , Representative sections from aged muscle stained with anti-Pax7, Ki67 and laminin. e , Percentage of BrdU + SCs (n=300 cells,4-6 animals/group). f , Percentage of Ki67 + /Pax7 + SCs/section (n=4-6 animals/group). g, h , RTqPCR; myod ( g ) p27 and spry1 ( h ) in SCs. (n=10,000 cells/condition performed in triplicate, 5 mice/group). Scale-bar; 20μm, *P
    Rat Anti Brdu, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2692 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad rat anti brdu
    Representative photomicrographs of Ki‐67 (A), <t>BrdU</t> (B), and <t>DCX</t> (C) immunostainings in the hippocampal dentate gyrus. Arrows point at Ki‐67 (A), BrdU (B), and DCX (C) labeled cells.
    Rat Anti Brdu, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 2111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Accurate Chemical & Scientific Corporation rat anti brdu
    p63 haploinsufficiency causes decreased SVZ NPCs and decreased adult-born olfactory neurons in 3-month-old mice. A , B , RT-PCR analysis for TAp63 and ΔNp63 mRNA ( A ) or mRNAs encoding the C-terminal splice variants p63α, β, and γ ( B ) in total RNA from the adult SVZ or from cultured adult SVZ neurospheres (NS). Postnatal day 1 skin (S) and cultured dermal skin-derived precursors (SKP) were used as controls because skin expresses mRNAs for all of the p63 isoforms, whereas SKPs express TAp63 but not ΔNp63 mRNA. The same samples were also analyzed for GAPDH mRNA. C , Micrographs of a coronal cortical section immunostained for p63 (red) and <t>NeuN</t> (green; the far right panel is the merge). The section was counterstained with Hoechst (blue). Scale bar, 50 μm. D , E , Micrographs of coronal sections through the SVZ immunostained for p63 (red) and nestin (green, D ) or GFAP (green, E ). Sections were counterstained with Hoechst 33258 (blue). LV denotes the lateral ventricle and arrows double-labeled cells. Scale bar, 25 μm. F , Micrographs of coronal sections through the p63 +/+ (p63WT) and p63 +/− (p63HET) olfactory bulb 30 d after <t>BrdU</t> injection, immunostained for BrdU (green) and NeuN (red). The boxed areas in the left column are shown at higher magnification in the right column. Arrows denote double-labeled cells. Scale bar, 200 μm. G , Quantification of the total number of BrdU-positive, NeuN-positive olfactory bulb neurons, as determined by analyzing 10 sections spanning the extent of the olfactory bulb similar to those in F . GCL, Granule cell layer. *** p
    Rat Anti Brdu, supplied by Accurate Chemical & Scientific Corporation, used in various techniques. Bioz Stars score: 93/100, based on 1180 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rat anti brdu antibody
    Mean ± standard deviation values of the number of <t>BrdU-DCX-positive</t> cells in the penumbra ischemic cortex (see the legend of Figure 1 for group abbreviations). The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P
    Rat Anti Brdu Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 873 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti brdu antibody bu1 75 icr1
    Spinal <t>NG2-positive</t> and Iba1-positive cells proliferate after PNS injury. (A) Experimental schedule and schematic diagram. <t>Bromodeoxyuridine</t> (50 mg/kg/day, i.p.) or vehicle was injected on days 0, 2, 4, and 6 after unilateral SNC and detected in the ipsilateral
    Anti Brdu Antibody Bu1 75 Icr1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Abcam rat monoclonal anti brdu
    Deletion of <t>Misu</t> increases dormancy of bulge stem cell. (A,B) Whole mount staining of wild-type and Misu −/− tail skin for LRC (green), keratin 14 (K14) (red) and DAPI (blue) after 4 months chase period. (C) Automated quantification of LRC in tail outer hair follicles in (A,B). (D) Frequency distributions of the intensity of the <t>BrdU-label</t> in LRC in (A,B). Error bars represent SEM (n = 10) from at least 3 independent experiments. (E,F) Flow cytometry for Itgα6 and CD34 in epidermis in telogen (P49) (E) and catagen (P40) (F). The percentage of double positive cells in each group is shown ± SEM (n = 3). The red lines in (E) indicate a cell population that increases in Misu −/− skin in telogen. (G,H) Epidermal cells sorted for CD34 +ve and low (L) and high (H) levels of Itgα6 were subjected to QPCR for bulge and hair germ markers as indicated (G). RNA levels were measured relative to GAPDH and presented as fold enrichment in Misu −/− mice versus wild-type controls. Error bars represent SEM (n = 3). (I) BrdU labelling and chasing regime to measure migration of LCR from bulge to hair germ at telogen at P47 (arrow). (J,K) Detection of LRC (green), Ki67 (red) and DAPI (blue) in high and low part of the bulge (straight line) and hair germ (HG) in hair follicles, indicated by dotted line, from wild-type (wt) (J) and Misu −/− mice (K). (L) Automated quantification of LRC in the whole hair follicle (total), high and low bulge region and the hair germ. Error bars indicate SEM (n = 5). (M) Percentages of hair follicles without LRC in the lower bulge region. Scale bars: 250 µm (A,B); 50 µm (J,K).
    Rat Monoclonal Anti Brdu, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 350 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioreclamationIVT rat anti brdu
    Activity-induced changes in the number of <t>BrdU-labeled</t> cells and new neurons in the dentate gyrus . (A,B) Confocal projections of the dentate gyrus (z-stack of 10 optical sections with 1.7-μm thickness). BrdU, Red; DCX, Blue. Scale bar, 25 μm in (A–D) Confocal image (optical section of 1-μm thickness) of STDSTD in (C) and RUNENR in (D) showing <t>NeuN,</t> Green; BrdU, Red, S100β, Blue; Inset: a, NeuN; b, BrdU; c, S100β. (D) Arrowheads in red indicating colocalization of BrdU and NeuN. Scale bar, 25 μm in (C,D) . (E–H) Brightfield images of BrdU-positive cells in the dentate gyrus. Scale bar (in E for E–H ), 100 μm.
    Rat Anti Brdu, supplied by BioreclamationIVT, used in various techniques. Bioz Stars score: 92/100, based on 234 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Accurate Chemical & Scientific Corporation rat monoclonal anti brdu
    S-phase labeling with <t>BrdU</t> <t>IHC</t> labeling for BrdU identified newborn cells in the subcortical white matter at 18 hours (Proliferation group, A and B) and four weeks (Survival group, C and D) after BrdU injection in GH/IGF-I replete (A and C) and deficient (B and D) rats. The cingulate cortex (Cg) and hippocampus (Hp) are identified in each section for orientation. Inset: medial CC at higher magnification. Scale bar = 250 µm, 50 µm (inset).
    Rat Monoclonal Anti Brdu, supplied by Accurate Chemical & Scientific Corporation, used in various techniques. Bioz Stars score: 91/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad rat monoclonal anti brdu
    X-gal-negative Newly Formed Cardiomyocytes Originate from LRCs, Which Consisted of CSCs or CPCs. (A) A CPC co-expressing Sca-1 and GATA4. In the two left panels, the arrowhead indicates a Sca-1 (green) and GATA4 (red) double-positive CPC surrounded by a basement membrane of cardiomyocytes (laminin in blue). The GATA4 signal co-localized with DAPI nuclear staining (blue) in the two right panels. Arrows indicate Sca-1-positive capillaries. Scale bar, 10 μm. (B) Partial overlap of LRCs and CPCs. The hearts of 10-week-old mice that were administered <t>BrdU</t> during the fetal period were immunohistochemically examined. Some anti-sarcomeric α-actinin (SA-actinin)-negative and BrdU-positive LRCs were Nkx2.5-positive (arrowhead, upper panels) or pGATA4-positive (arrowhead, lower panels). Some Nkx2.5-positive (arrow, upper panels) or pGATA4-positive (arrow, lower panels) cardiomyocytes retained BrdU. Nuclei were stained with DAPI. Scale bar, 10 μm. (C) Partial overlap of LRCs and CSCs. Cardiac side population cells (CSPs) were isolated from the hearts of 10-week-old mice that had been administered BrdU during the fetal period. BrdU-positive and multi-drug-resistant protein 1(MDR1)-positive CSPs were identified (arrowheads). Scale bar, 10 μm. (D) X-gal-negative newly formed cardiomyocytes originate from LRCs. A white arrowhead indicates SA-actinin-positive and X-gal-negative LRC-derived newly formed cardiomyocytes. An arrow indicates SA-actinin- and X-gal-positive pre-existing cardiomyocytes. Scale bar, 10 μm. Note that the nuclei of both cell types retained BrdU because of their quiescence after birth. Two yellow arrowheads indicate SA-actinin-positive and X-gal-negative cardiomyocytes, the ancestral CSCs or CPCs of which circumvented the BrdU labeling because of their quiescence. (E) Cre-mediated recombination does not occur in CSPs after tamoxifen treatment. X-gal staining of the cardiomyocyte fraction (right) and CSP cell fraction (left) from a tamoxifen-treated mouse. Arrowheads indicate X-gal-negative CSPs. An arrow indicates X-gal-positive cardiomyocytes. β-galactosidase mRNA expression in cardiomyocyte suspension and sorted CSP fraction from tamoxifen-treated and -untreated mice. Scale bar, 10 μm.
    Rat Monoclonal Anti Brdu, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals rat anti brdu
    Fluorescence microscopy of neuronal and microglial cells derived from dividing progenitors. Panel A is a confocal photomicrograph that shows <t>NeuN+</t> (red) cells in the cortex of a control mouse brain for the striatal line. Panel B is a confocal photomicrograph that shows a <t>BrdU+</t> (green) cell in the same field. Panel C represents the merging of panels A and B , and indicates that the cell (arrow) is NeuN+/BrdU+ (yellow) and therefore represents a neuron derived from a dividing progenitor cell. Scale bar in panels A , B and C represents 12 μm. Panel D is a confocal photomicrograph that shows Iba1+ (red) cells in the striatum of a mutant mouse brain from the striatal line. Panel E is a confocal photomicrograph that shows BrdU+ (green) cells in the same field. Panel F represents the merging of panels D and E and indicates that one cell (arrow) is Iba1+/BrdU+ (yellow) and therefore represents a microglial cell derived from a cell that has undergone cell division. Scale bar in panels D , E and F represents 15 μm.
    Rat Anti Brdu, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 97/100, based on 147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rat anti brdu
    Depletion of slowly dividing neural stem cells and impairment of regeneration after cytosine-β-D-arabinofuranoside (AraC) treatment in <t>Hes1/3/5/Hey1</t> mutant adult mouse brains. ( A ) Experimental design. ( B – E ) Dividing progenitors ( t = 0; B , C ) and slowly dividing cells, ones that retained labeling even after 12 d ( t = 12; D , E ), in the SVZ were labeled with <t>BrdU</t> in control ( B , D ) and Hes1/3/5/Hey1 mutant ( C , E ) mice. ( F , G ) Quantification of the number of BrdU-incorporating cells in the SVZ at t = 0 and t = 12 after BrdU administration for 14 consecutive days. WT-control ( Hes1 +/+ ;Hes3 +/+ ;Hes5 +/+ ;Hey1 +/+ ) mice are shown as a reference. ( H ) Experimental design. ( I – P ) Coronal sections of the SVZ showing BrdU-incorporating cells soon after the removal of the osmotic pump ( t = 0; I , J , M , N ) and 10 d after the pump removal ( t = 10; K , L , O , P ) in AraC-treated control ( I – L ) and AraC-treated Hes1/3/5/Hey1 mutant ( M – P ) mice. ( Q ) Quantification of the number of BrdU-incorporating cells in the SVZ. BrdU was administered for 4 h before sacrifice. (*) P
    Rat Anti Brdu, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rat anti brdu monoclonal antibody
    CM-specific overexpression of dn-c-kit resulted in CM hypertrophy, leading to a thicker LV wall, a smaller LV cavity and higher ejection fraction. ( a ) Expression of Kit (c-Kit) mRNA in male P196 dn-c-kit-Tg (Tg; n = 11) and NTL (n = 6) hearts; differences analysed by Student’s t -test. (b , c ) Examples (white arrowheads) of <t>BrdU</t> + <t>/cTnT</t> + ( b ) or H3P + /cTnT + ( c ) CMs isolated from P121 Tg ( b , right panel, and c ) or NTL ( b , left panel) hearts after 9 days of BrdU (10 mg/kg/day) infusion by osmotic mini-pump. Scale bar: 50 μm. ( d ) Left panels, representative photographs, and right panel, quantitation of surface area showing hypertrophy of isolated CMs from P121 Tg (682 CMs analysed from n = 3 mice) relative to NTL hearts (1015 CMs analysed from n = 3 mice); differences analysed by Student’s t -test. Scale bar: 100 μm. ( e ) Abundance of Myh6 (encoding α-MHC) and Acta1 (α-skeletal actin) mRNAs was similar, while abundance of other mRNAs ( Nppa (ANP), Nppb (BNP), Myh 7 (β-MHC)) and the Myh 7 :Myh6 (β-:α-MHC) ratio were significantly increased in male P196 dn-c-kit-Tg relative to NTL hearts (n = 7); differences analysed by Student’s t -test. ( f ) Increased LV wall thickness to chamber radius ( h/r ) ratio at end-diastole in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test. ( g ) Increased ejection fraction in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test.
    Rat Anti Brdu Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson rat anti brdu
    p107 regulates the neural precursor population through the repression of Hes1. (a) Adult mice received intraperitoneal injections of <t>BrdU</t> to label proliferating cells over a 10-h period. BrdU-positive cells were counted in every 10th section through the forebrains of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 4), and p107 −/− ( n = 5) mice. (b) Proliferating progenitors in E10.5 brains were identified by <t>PH3</t> immunohistochemistry. PH3-positive cells were counted in three representative sections of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 3), and p107 −/− ( n = 3) brains. (c) A 2-h BrdU pulse labeled proliferating progenitors in E13.5 brains. BrdU-positive cells were counted in four representative regions of the brain in wild-type ( n = 4), Hes1 +/− :p107 −/− ( n = 4), Hes1 +/− ( n = 4), and p107 −/− ( n = 3) embryos. Note that loss of a single Hes1 allele restored the numbers of progenitor cells to wild-type levels in p107 −/− mice at embryonic and adult ages. Means were statistically analyzed by one-way analysis of variance followed by Tukey's individual comparison of the means. Error bars represent SEM. *, P
    Rat Anti Brdu, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Adult hippocampus and OB of D2 KO mice deficient in neurogenesis. (a–d) Extensive labeling in hippocampal DG in controls: wild-type (WT) from D1 KO line (c), WT from D2 KO line (d) and D1 KO mouse (a). Lack of labeling in DG of D2 KO hippocampus (b). (e–h) Similar phenomenon in OB. (i and j) Colocalization (arrows) of BrdU (red) and NeuN (green) in WT hippocampus. (k and l) BrdU-positive nuclei in D2 KO hippocampus never colocalize with NeuN. (o) Similar colocalization (arrowheads indicating green rings) of BrdU (red) and Tuj-1 (green) with no colocalization in D2 KO (p). (m and n) Colabeling of BrdU with glial fibrillary acidic protein (GFAP, green) in D2 KO hippocampus. Confocal images: single-plane (i–l and o and p) and composed (m and n). Bars: (a–h) 200 μm; (i–l, m and n, and o and p) 20 μm.

    Journal: The Journal of Cell Biology

    Article Title: The critical role of cyclin D2 in adult neurogenesis

    doi: 10.1083/jcb.200404181

    Figure Lengend Snippet: Adult hippocampus and OB of D2 KO mice deficient in neurogenesis. (a–d) Extensive labeling in hippocampal DG in controls: wild-type (WT) from D1 KO line (c), WT from D2 KO line (d) and D1 KO mouse (a). Lack of labeling in DG of D2 KO hippocampus (b). (e–h) Similar phenomenon in OB. (i and j) Colocalization (arrows) of BrdU (red) and NeuN (green) in WT hippocampus. (k and l) BrdU-positive nuclei in D2 KO hippocampus never colocalize with NeuN. (o) Similar colocalization (arrowheads indicating green rings) of BrdU (red) and Tuj-1 (green) with no colocalization in D2 KO (p). (m and n) Colabeling of BrdU with glial fibrillary acidic protein (GFAP, green) in D2 KO hippocampus. Confocal images: single-plane (i–l and o and p) and composed (m and n). Bars: (a–h) 200 μm; (i–l, m and n, and o and p) 20 μm.

    Article Snippet: For double labeling, sections were incubated overnight with pooled primary antibodies: rat-anti-BrdU (1:100; Accurate) together with mouse anti-NeuN (1:500; Chemicon) or mouse anti–Tuj-1 (1:100; R & D Systems), or mouse anti-GFAP (1:500; Chemicon) followed by goat anti–rat Cy3 (Chemicon; red) and goat anti–mouse (Alexa-Fluor 488; Molecular Probes; green).

    Techniques: Mouse Assay, Labeling

    Aged satellite cells cycle more frequently during homeostasis a , Doxycycline (Dox) loading and chase strategy of TetO-H2B-GFP mice and FACS plots for No chase and 84-week chase. SC pool has LRC and nonLRC subsets. b , Strategy and FACS plots for aged (Ag) or adult (Ad) no chase and 12-week chase SCs. Vehicle treated SCs (Neg) (n=10,000 cells,6 animals/group). c , BrdU feeding and representative images of SCs stained with anti-Pax7, BrdU and Dapi. d , Representative sections from aged muscle stained with anti-Pax7, Ki67 and laminin. e , Percentage of BrdU + SCs (n=300 cells,4-6 animals/group). f , Percentage of Ki67 + /Pax7 + SCs/section (n=4-6 animals/group). g, h , RTqPCR; myod ( g ) p27 and spry1 ( h ) in SCs. (n=10,000 cells/condition performed in triplicate, 5 mice/group). Scale-bar; 20μm, *P

    Journal: Nature

    Article Title: The aged niche disrupts muscle stem cell quiescence

    doi: 10.1038/nature11438

    Figure Lengend Snippet: Aged satellite cells cycle more frequently during homeostasis a , Doxycycline (Dox) loading and chase strategy of TetO-H2B-GFP mice and FACS plots for No chase and 84-week chase. SC pool has LRC and nonLRC subsets. b , Strategy and FACS plots for aged (Ag) or adult (Ad) no chase and 12-week chase SCs. Vehicle treated SCs (Neg) (n=10,000 cells,6 animals/group). c , BrdU feeding and representative images of SCs stained with anti-Pax7, BrdU and Dapi. d , Representative sections from aged muscle stained with anti-Pax7, Ki67 and laminin. e , Percentage of BrdU + SCs (n=300 cells,4-6 animals/group). f , Percentage of Ki67 + /Pax7 + SCs/section (n=4-6 animals/group). g, h , RTqPCR; myod ( g ) p27 and spry1 ( h ) in SCs. (n=10,000 cells/condition performed in triplicate, 5 mice/group). Scale-bar; 20μm, *P

    Article Snippet: REAGENTS AND ANTIBODIES The source and concentration of antibodies: Rat anti-BrdU (1/500, Abcam), rabbit anti-ki67 (1/500, Abcam) mouse anti-Pax7 (1/100, DSHB), rabbit anti-Myogenin (1/250, Santa Cruz), Cleaved Caspase-3 (1/500, Cell Signaling Technologies), chick anti-Laminin (1/5000, Abcam), VCAM (1/100), mouse anti-Integrin-α7 (1/200, MBL), CD31-PE, CD45-PE (1/200, BD) and rabbit anti-FGF2 (1/500 Abcam).

    Techniques: Mouse Assay, FACS, Staining

    Aged satellite cells are sensitive to acute changes of FGF signalling a-c , Strategy to delete Spry1 in SCs of adult (Ad) and aged (Ag) mice prior to injection of SU5402-coated beads ( b ) number of Pax7 + SCs/single muscle fibre (20-30 single muscle fibers/animal, n=4-6 mice/group) ( c ) percent Ki67 + /Pax7 + SCs per 10μm cross-section 10 days post Tmx (n= 4-6 animals/group). d , Strategy to label and chase BrdU in SCs from Ad and Ag Ctrl or Spry1null mice 6 weeks post Tmx injection and Percent BrdU + /LRC SCs after chase. e , Strategy for BrdU incorporation in Tmx-treated Ctrl and Spry1OX aged mice and Percent BrdU + /SCs after chase. For panels d and e , n=1000 cells /condition, 4-6 mice/group. *P

    Journal: Nature

    Article Title: The aged niche disrupts muscle stem cell quiescence

    doi: 10.1038/nature11438

    Figure Lengend Snippet: Aged satellite cells are sensitive to acute changes of FGF signalling a-c , Strategy to delete Spry1 in SCs of adult (Ad) and aged (Ag) mice prior to injection of SU5402-coated beads ( b ) number of Pax7 + SCs/single muscle fibre (20-30 single muscle fibers/animal, n=4-6 mice/group) ( c ) percent Ki67 + /Pax7 + SCs per 10μm cross-section 10 days post Tmx (n= 4-6 animals/group). d , Strategy to label and chase BrdU in SCs from Ad and Ag Ctrl or Spry1null mice 6 weeks post Tmx injection and Percent BrdU + /LRC SCs after chase. e , Strategy for BrdU incorporation in Tmx-treated Ctrl and Spry1OX aged mice and Percent BrdU + /SCs after chase. For panels d and e , n=1000 cells /condition, 4-6 mice/group. *P

    Article Snippet: REAGENTS AND ANTIBODIES The source and concentration of antibodies: Rat anti-BrdU (1/500, Abcam), rabbit anti-ki67 (1/500, Abcam) mouse anti-Pax7 (1/100, DSHB), rabbit anti-Myogenin (1/250, Santa Cruz), Cleaved Caspase-3 (1/500, Cell Signaling Technologies), chick anti-Laminin (1/5000, Abcam), VCAM (1/100), mouse anti-Integrin-α7 (1/200, MBL), CD31-PE, CD45-PE (1/200, BD) and rabbit anti-FGF2 (1/500 Abcam).

    Techniques: Mouse Assay, Injection, BrdU Incorporation Assay

    Representative photomicrographs of Ki‐67 (A), BrdU (B), and DCX (C) immunostainings in the hippocampal dentate gyrus. Arrows point at Ki‐67 (A), BrdU (B), and DCX (C) labeled cells.

    Journal: CNS Neuroscience & Therapeutics

    Article Title: The Novel Antidepressant Agomelatine Normalizes Hippocampal Neuronal Activity and Promotes Neurogenesis in Chronically Stressed Rats

    doi: 10.1111/j.1755-5949.2009.00125.x

    Figure Lengend Snippet: Representative photomicrographs of Ki‐67 (A), BrdU (B), and DCX (C) immunostainings in the hippocampal dentate gyrus. Arrows point at Ki‐67 (A), BrdU (B), and DCX (C) labeled cells.

    Article Snippet: Brain sections for all four immunostainings were preincubated in 3% normal serum and 0.1% TritonX‐100, and then incubated with one of the following antibodies for 60 h at 4°C: primary mouse‐anti‐Ki‐67 (1:200; Monosan, Uden, The Netherlands), goat‐anti‐DCX (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rat‐anti‐BrdU (1:800; Serotec, Oxford, UK) or rabbit‐anti‐c‐Fos (1:10000; Calbiochem, Darmstadt, Germany).

    Techniques: Labeling

    Results of the histological analysis of adult hippocampal neurogenesis in brain sections from WT, TgN3 WT and TgN3 R169C mice after 28 days ( a and c ) or 6 months ( b and d ) under standard (STD), running wheel (RUN) or environmentally enriched (ENR) cage conditions. The percentage of BrdU+/S100β+ ( a and b ) and BrdU+/NeuN+ cells ( c and d ) of all BrdU+ cells was determined to assess effects on the differentiation of BrdU+ cells to astrocytes and neurons. The percentage of BrdU+/S100β+ cells in TgN3 R169C is increased in older mice independent of RUN or ENR ( b ). Data are expressed as mean ± S.E.M.

    Journal: Scientific Reports

    Article Title: Stimulation of adult hippocampal neurogenesis by physical exercise and enriched environment is disturbed in a CADASIL mouse model

    doi: 10.1038/srep45372

    Figure Lengend Snippet: Results of the histological analysis of adult hippocampal neurogenesis in brain sections from WT, TgN3 WT and TgN3 R169C mice after 28 days ( a and c ) or 6 months ( b and d ) under standard (STD), running wheel (RUN) or environmentally enriched (ENR) cage conditions. The percentage of BrdU+/S100β+ ( a and b ) and BrdU+/NeuN+ cells ( c and d ) of all BrdU+ cells was determined to assess effects on the differentiation of BrdU+ cells to astrocytes and neurons. The percentage of BrdU+/S100β+ cells in TgN3 R169C is increased in older mice independent of RUN or ENR ( b ). Data are expressed as mean ± S.E.M.

    Article Snippet: Therefore, a one-in-six series of free-floating brain sections of each animal was pretreated with HCl, followed by an overnight incubation at 4 °C with primary rat anti-BrdU antibody (AbD serotec, 1:500), mouse anti-NeuN (Millipore, 1:1000) and rabbit anti-S100β (Abcam, 1:150).

    Techniques: Mouse Assay

    Results of the histological analysis of adult hippocampal neurogenesis in brain sections from WT, TgN3 WT and TgN3 R169C mice after 28 days ( a , c and e ) or 6 months ( b , d and f ) under standard (STD), running wheel (RUN) or environmentally enriched (ENR) cage conditions. The absolute number of BrdU+ ( a and b ), BrdU+/S100β+ ( c and d ) and BrdU+/NeuN+ cells ( e and f ) was quantified to determine the survival rate of proliferating cells, new astrocytic and new neuronal cells. New neuron survival is reduced in older ( f ) but not younger TgN3 WT mice ( e ). Neurogenic stimulation by RUN or ENR failed in both TgN3 WT and TgN3 R169C independent of the duration ( e and f ). Data are expressed as mean ± S.E.M. *p

    Journal: Scientific Reports

    Article Title: Stimulation of adult hippocampal neurogenesis by physical exercise and enriched environment is disturbed in a CADASIL mouse model

    doi: 10.1038/srep45372

    Figure Lengend Snippet: Results of the histological analysis of adult hippocampal neurogenesis in brain sections from WT, TgN3 WT and TgN3 R169C mice after 28 days ( a , c and e ) or 6 months ( b , d and f ) under standard (STD), running wheel (RUN) or environmentally enriched (ENR) cage conditions. The absolute number of BrdU+ ( a and b ), BrdU+/S100β+ ( c and d ) and BrdU+/NeuN+ cells ( e and f ) was quantified to determine the survival rate of proliferating cells, new astrocytic and new neuronal cells. New neuron survival is reduced in older ( f ) but not younger TgN3 WT mice ( e ). Neurogenic stimulation by RUN or ENR failed in both TgN3 WT and TgN3 R169C independent of the duration ( e and f ). Data are expressed as mean ± S.E.M. *p

    Article Snippet: Therefore, a one-in-six series of free-floating brain sections of each animal was pretreated with HCl, followed by an overnight incubation at 4 °C with primary rat anti-BrdU antibody (AbD serotec, 1:500), mouse anti-NeuN (Millipore, 1:1000) and rabbit anti-S100β (Abcam, 1:150).

    Techniques: Mouse Assay

    Representative confocal images of the triple fluorescent staining of the DG of two different mice ( a – d) : TgN3 WT ENR 6 months; ( e – h ) TgN3 WT STD 28 days). Arrows point to BrdU+ cell nuclei (red, a and e ), NeuN+ cell nuclei (cyan, b and f ), S100β+ cells (green, c and g ), two BrdU+/NeuN+ cell nuclei ( d ) and a BrdU+/S100β+ cell ( h ). Scale bar = 50 μm.

    Journal: Scientific Reports

    Article Title: Stimulation of adult hippocampal neurogenesis by physical exercise and enriched environment is disturbed in a CADASIL mouse model

    doi: 10.1038/srep45372

    Figure Lengend Snippet: Representative confocal images of the triple fluorescent staining of the DG of two different mice ( a – d) : TgN3 WT ENR 6 months; ( e – h ) TgN3 WT STD 28 days). Arrows point to BrdU+ cell nuclei (red, a and e ), NeuN+ cell nuclei (cyan, b and f ), S100β+ cells (green, c and g ), two BrdU+/NeuN+ cell nuclei ( d ) and a BrdU+/S100β+ cell ( h ). Scale bar = 50 μm.

    Article Snippet: Therefore, a one-in-six series of free-floating brain sections of each animal was pretreated with HCl, followed by an overnight incubation at 4 °C with primary rat anti-BrdU antibody (AbD serotec, 1:500), mouse anti-NeuN (Millipore, 1:1000) and rabbit anti-S100β (Abcam, 1:150).

    Techniques: Staining, Mouse Assay

    KA treatment enhances NSC proliferation and neuroblast migration in the dentate gyrus (DG). (A) Representative image of a section of the DG, showing the division into granular zone (GZ), subgranular zone (SGZ) and hilus. EdU in green and nuclei, stained for Hoechst 33342, in gray. Scale bar: 50 μm. (B) WT and hCAST mice were treated with either SAL or KA and were given BrdU on the day before sacrifice, to assess cell proliferation. Representative images from hippocampal brain sections for each group (left panels), showing BrdU-positive cells in white. Scale bar: 200 μm. BrdU-positive cells were counted in the SGZ of five central coronal sections of the hippocampal region for each animal (right panel). (C) WT and hCAST mice were treated with either SAL or KA and sacrificed after 14 days, to assess cell migration. Representative images from hippocampal brain sections for each group (left panels), showing migrating neuroblasts, labeled for doublecortin (DCX), in white. Scale bar: 100 μm. Percentage of DCX-positive area was determined in the DG of five central coronal sections of the hippocampal region for each animal (right panel). Data are presented as means ± SEM of 9–14 animals per group (for cell proliferation) and 6–9 animals per group (for cell migration). Statistical significance was determined using the Kruskal-Wallis test (Dunn’s post-test), * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Calpastatin Overexpression Preserves Cognitive Function Following Seizures, While Maintaining Post-Injury Neurogenesis

    doi: 10.3389/fnmol.2017.00060

    Figure Lengend Snippet: KA treatment enhances NSC proliferation and neuroblast migration in the dentate gyrus (DG). (A) Representative image of a section of the DG, showing the division into granular zone (GZ), subgranular zone (SGZ) and hilus. EdU in green and nuclei, stained for Hoechst 33342, in gray. Scale bar: 50 μm. (B) WT and hCAST mice were treated with either SAL or KA and were given BrdU on the day before sacrifice, to assess cell proliferation. Representative images from hippocampal brain sections for each group (left panels), showing BrdU-positive cells in white. Scale bar: 200 μm. BrdU-positive cells were counted in the SGZ of five central coronal sections of the hippocampal region for each animal (right panel). (C) WT and hCAST mice were treated with either SAL or KA and sacrificed after 14 days, to assess cell migration. Representative images from hippocampal brain sections for each group (left panels), showing migrating neuroblasts, labeled for doublecortin (DCX), in white. Scale bar: 100 μm. Percentage of DCX-positive area was determined in the DG of five central coronal sections of the hippocampal region for each animal (right panel). Data are presented as means ± SEM of 9–14 animals per group (for cell proliferation) and 6–9 animals per group (for cell migration). Statistical significance was determined using the Kruskal-Wallis test (Dunn’s post-test), * p

    Article Snippet: After blocking, the sections were kept with the primary antibodies—rat anti-BrdU 1:50 (AbD Serotec, Oxford, UK), goat anti-DCX 1:400 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), mouse anti-NeuN 1:200 (Merck Millipore, Billerica, MA, USA)—for 48 h, at 4°C.

    Techniques: Migration, Staining, Mouse Assay, Labeling

    p63 haploinsufficiency causes decreased SVZ NPCs and decreased adult-born olfactory neurons in 3-month-old mice. A , B , RT-PCR analysis for TAp63 and ΔNp63 mRNA ( A ) or mRNAs encoding the C-terminal splice variants p63α, β, and γ ( B ) in total RNA from the adult SVZ or from cultured adult SVZ neurospheres (NS). Postnatal day 1 skin (S) and cultured dermal skin-derived precursors (SKP) were used as controls because skin expresses mRNAs for all of the p63 isoforms, whereas SKPs express TAp63 but not ΔNp63 mRNA. The same samples were also analyzed for GAPDH mRNA. C , Micrographs of a coronal cortical section immunostained for p63 (red) and NeuN (green; the far right panel is the merge). The section was counterstained with Hoechst (blue). Scale bar, 50 μm. D , E , Micrographs of coronal sections through the SVZ immunostained for p63 (red) and nestin (green, D ) or GFAP (green, E ). Sections were counterstained with Hoechst 33258 (blue). LV denotes the lateral ventricle and arrows double-labeled cells. Scale bar, 25 μm. F , Micrographs of coronal sections through the p63 +/+ (p63WT) and p63 +/− (p63HET) olfactory bulb 30 d after BrdU injection, immunostained for BrdU (green) and NeuN (red). The boxed areas in the left column are shown at higher magnification in the right column. Arrows denote double-labeled cells. Scale bar, 200 μm. G , Quantification of the total number of BrdU-positive, NeuN-positive olfactory bulb neurons, as determined by analyzing 10 sections spanning the extent of the olfactory bulb similar to those in F . GCL, Granule cell layer. *** p

    Journal: The Journal of Neuroscience

    Article Title: p63 Regulates Adult Neural Precursor and Newly Born Neuron Survival to Control Hippocampal-Dependent Behavior

    doi: 10.1523/JNEUROSCI.1251-13.2013

    Figure Lengend Snippet: p63 haploinsufficiency causes decreased SVZ NPCs and decreased adult-born olfactory neurons in 3-month-old mice. A , B , RT-PCR analysis for TAp63 and ΔNp63 mRNA ( A ) or mRNAs encoding the C-terminal splice variants p63α, β, and γ ( B ) in total RNA from the adult SVZ or from cultured adult SVZ neurospheres (NS). Postnatal day 1 skin (S) and cultured dermal skin-derived precursors (SKP) were used as controls because skin expresses mRNAs for all of the p63 isoforms, whereas SKPs express TAp63 but not ΔNp63 mRNA. The same samples were also analyzed for GAPDH mRNA. C , Micrographs of a coronal cortical section immunostained for p63 (red) and NeuN (green; the far right panel is the merge). The section was counterstained with Hoechst (blue). Scale bar, 50 μm. D , E , Micrographs of coronal sections through the SVZ immunostained for p63 (red) and nestin (green, D ) or GFAP (green, E ). Sections were counterstained with Hoechst 33258 (blue). LV denotes the lateral ventricle and arrows double-labeled cells. Scale bar, 25 μm. F , Micrographs of coronal sections through the p63 +/+ (p63WT) and p63 +/− (p63HET) olfactory bulb 30 d after BrdU injection, immunostained for BrdU (green) and NeuN (red). The boxed areas in the left column are shown at higher magnification in the right column. Arrows denote double-labeled cells. Scale bar, 200 μm. G , Quantification of the total number of BrdU-positive, NeuN-positive olfactory bulb neurons, as determined by analyzing 10 sections spanning the extent of the olfactory bulb similar to those in F . GCL, Granule cell layer. *** p

    Article Snippet: For immunostaining, the primary antibodies used were mouse anti-p63 (1:200), mouse anti-NeuN (1:500; Millipore Bioscience Research Reagents), Alexa Fluor 488-conjugated mouse anti-NeuN (1:100; Millipore), rat anti-BrdU (1:200; Accurate Chemical), rabbit anti-Sox2 (1:200; Cell Signaling Technology), goat anti-Sox2 (1:50; Santa Cruz Biotechnology), rabbit anti-GFAP (1:1000; Dako), goat anti-DCX (1:200; Santa Cruz Biotechnology), rabbit anti-CC3 (1:500; Cell Signaling Technology), chicken anti-nestin (1:1000; Aves Labs), and chicken anti-GFP (1:1000; Millipore).

    Techniques: Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Derivative Assay, Labeling, Injection

    Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the penumbra ischemic cortex (see the legend of Figure 1 for group abbreviations). The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Journal: Mediators of Inflammation

    Article Title: Etanercept Attenuates Traumatic Brain Injury in Rats by Reducing Brain TNF-α Contents and by Stimulating Newly Formed Neurogenesis

    doi: 10.1155/2013/620837

    Figure Lengend Snippet: Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the penumbra ischemic cortex (see the legend of Figure 1 for group abbreviations). The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Article Snippet: The antibodies therein were, sequentially, rabbit anti-DCX antibody (Cell Signaling Technology, 1 : 200), rat anti-BrdU antibody (Abcam, 1 : 200), goat anti-rabbit IgG-H & L antibody (Abcam, 1 : 400), and goat anti-rat IgG antibody (Abcam, 1 : 400).

    Techniques: Standard Deviation

    Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the ischemic hippocampus. (See the legend of Figure 1 for group abbreviations). The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Journal: Mediators of Inflammation

    Article Title: Etanercept Attenuates Traumatic Brain Injury in Rats by Reducing Brain TNF-α Contents and by Stimulating Newly Formed Neurogenesis

    doi: 10.1155/2013/620837

    Figure Lengend Snippet: Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the ischemic hippocampus. (See the legend of Figure 1 for group abbreviations). The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Article Snippet: The antibodies therein were, sequentially, rabbit anti-DCX antibody (Cell Signaling Technology, 1 : 200), rat anti-BrdU antibody (Abcam, 1 : 200), goat anti-rabbit IgG-H & L antibody (Abcam, 1 : 400), and goat anti-rat IgG antibody (Abcam, 1 : 400).

    Techniques: Standard Deviation

    Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the core ischemic cortex. Sham-TBI (white column): rats given a sham traumatic brain injury (TBI) operation; TBI + V (lined column): TBI rats treated with vehicle solution; TBI + E (crossed column): TBI rats treated with etanercept solution. The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Journal: Mediators of Inflammation

    Article Title: Etanercept Attenuates Traumatic Brain Injury in Rats by Reducing Brain TNF-α Contents and by Stimulating Newly Formed Neurogenesis

    doi: 10.1155/2013/620837

    Figure Lengend Snippet: Mean ± standard deviation values of the number of BrdU-DCX-positive cells in the core ischemic cortex. Sham-TBI (white column): rats given a sham traumatic brain injury (TBI) operation; TBI + V (lined column): TBI rats treated with vehicle solution; TBI + E (crossed column): TBI rats treated with etanercept solution. The data were obtained 7 days after TBI or sham-TBI operation ( n = 8). * P

    Article Snippet: The antibodies therein were, sequentially, rabbit anti-DCX antibody (Cell Signaling Technology, 1 : 200), rat anti-BrdU antibody (Abcam, 1 : 200), goat anti-rabbit IgG-H & L antibody (Abcam, 1 : 400), and goat anti-rat IgG antibody (Abcam, 1 : 400).

    Techniques: Standard Deviation

    Detection of BrdU-labeled nuclei in tissue sections Confocal immunofluorescence images show BrdU labeled nuclei (red) in heart sections from mice injected with BrdU and total nuclei stained with DAPI (blue). Overlaid image shows overlap (arrowheads) of BrdU stained nuclei with DAPI. Scale bar = 78 μm.

    Journal: Current protocols in molecular biology

    Article Title: Detection of markers of cell proliferation by immunofluorescent staining and microscopy imaging in paraffin-embedded tissue sections

    doi: 10.1002/cpmb.13

    Figure Lengend Snippet: Detection of BrdU-labeled nuclei in tissue sections Confocal immunofluorescence images show BrdU labeled nuclei (red) in heart sections from mice injected with BrdU and total nuclei stained with DAPI (blue). Overlaid image shows overlap (arrowheads) of BrdU stained nuclei with DAPI. Scale bar = 78 μm.

    Article Snippet: While the specific nuclear BrdU staining can be distinguished from the non-specific staining by co-localisation of BrdU signal with nuclear DAPI staining , if non-specific staining is significant, rat monoclonal anti-BrdU antibodies (Clone Bu1/75 ICR, Abcam) are recommended.

    Techniques: Labeling, Immunofluorescence, Mouse Assay, Injection, Staining

    Cell proliferation and apoptosis in MMTV-PyMT; Apc Min/+ and MMTV-PyMT; Apc +/+ cells after treatment with paclitaxel, cisplatin and doxorubicin. a Cell proliferation as measured by BrdU incorporation after chemotherapuetic treatment. MMTV-PyMT; Apc Min/+ cells showed a modest decrease in proliferation after treatment with cisplatin and doxorubicin compared to MMTV-PyMT; Apc +/+ cells. b Apoptosis as measured by cleaved caspase 3 immunofluorescence (IF). The percentage of apoptosis was lower in cisplatin and doxorubicin treated MMTV-PyMT; Apc Min/+ compared to MMTV-PyMT; Apc +/+ cells while paclitaxel treatment did not affect apoptosis levels. c Representative images of cleaved caspase 3 (CC3) IF. The scale bar is equal to 200 microns. White arrows are representative cleaved caspase 3 positive cells. Data are shown as the means ± SEM from 3 independent experiments; *P

    Journal: BMC Cancer

    Article Title: APC selectively mediates response to chemotherapeutic agents in breast cancer

    doi: 10.1186/s12885-015-1456-x

    Figure Lengend Snippet: Cell proliferation and apoptosis in MMTV-PyMT; Apc Min/+ and MMTV-PyMT; Apc +/+ cells after treatment with paclitaxel, cisplatin and doxorubicin. a Cell proliferation as measured by BrdU incorporation after chemotherapuetic treatment. MMTV-PyMT; Apc Min/+ cells showed a modest decrease in proliferation after treatment with cisplatin and doxorubicin compared to MMTV-PyMT; Apc +/+ cells. b Apoptosis as measured by cleaved caspase 3 immunofluorescence (IF). The percentage of apoptosis was lower in cisplatin and doxorubicin treated MMTV-PyMT; Apc Min/+ compared to MMTV-PyMT; Apc +/+ cells while paclitaxel treatment did not affect apoptosis levels. c Representative images of cleaved caspase 3 (CC3) IF. The scale bar is equal to 200 microns. White arrows are representative cleaved caspase 3 positive cells. Data are shown as the means ± SEM from 3 independent experiments; *P

    Article Snippet: Cells were incubated with primary antibodies: anti-BrdU rat monoclonal antibody (1:300, Abcam, Cambridge, MA), anti-cleaved caspase 3 rabbit monoclonal antibody (1:400, Cell Signaling Technology, Danvers, MA) or anti-APC (1:400, a gift from K. Neufeld, University of Kansas) for 1 h at 37 °C.

    Techniques: BrdU Incorporation Assay, Immunofluorescence

    APC knockdown in MDA-MB-157 cells impacts response to paclitaxel and cisplatin. a Quantitative RT-PCR in MDA-MB-157 cells and shAPC constructs shows decreased level of APC in cells infected with the shAPC constructs. b Representative APC immunofluorescence images showing that APC knockdown cells have less APC protein compared to the MDA-MB-157 parent line. c Cell proliferation as measured by BrdU incorporation did not differ between the three cell lines after treatment with cisplatin, doxorubicin or paclitaxel. d Apoptosis as measured by cleaved caspase 3 IF. The percentage of apoptosis was lower in paclitaxel treated shAPC 1 and cisplatin treated shAPC 2 cells compared to MDA-MB-157 control cells. Doxorubicin treatment had no effect on rates of apoptosis. e Representative images of CC3 IF. Although there are a similar number of positive cells in many of the images, there are fewer total cells in those images representing treatments with a higher percent of apoptosis. The scale bar is equal to 100 microns (E) and 20 microns (B). Data are shown as the means ± SEM from 3 independent experiments; *P

    Journal: BMC Cancer

    Article Title: APC selectively mediates response to chemotherapeutic agents in breast cancer

    doi: 10.1186/s12885-015-1456-x

    Figure Lengend Snippet: APC knockdown in MDA-MB-157 cells impacts response to paclitaxel and cisplatin. a Quantitative RT-PCR in MDA-MB-157 cells and shAPC constructs shows decreased level of APC in cells infected with the shAPC constructs. b Representative APC immunofluorescence images showing that APC knockdown cells have less APC protein compared to the MDA-MB-157 parent line. c Cell proliferation as measured by BrdU incorporation did not differ between the three cell lines after treatment with cisplatin, doxorubicin or paclitaxel. d Apoptosis as measured by cleaved caspase 3 IF. The percentage of apoptosis was lower in paclitaxel treated shAPC 1 and cisplatin treated shAPC 2 cells compared to MDA-MB-157 control cells. Doxorubicin treatment had no effect on rates of apoptosis. e Representative images of CC3 IF. Although there are a similar number of positive cells in many of the images, there are fewer total cells in those images representing treatments with a higher percent of apoptosis. The scale bar is equal to 100 microns (E) and 20 microns (B). Data are shown as the means ± SEM from 3 independent experiments; *P

    Article Snippet: Cells were incubated with primary antibodies: anti-BrdU rat monoclonal antibody (1:300, Abcam, Cambridge, MA), anti-cleaved caspase 3 rabbit monoclonal antibody (1:400, Cell Signaling Technology, Danvers, MA) or anti-APC (1:400, a gift from K. Neufeld, University of Kansas) for 1 h at 37 °C.

    Techniques: Multiple Displacement Amplification, Quantitative RT-PCR, Construct, Infection, Immunofluorescence, BrdU Incorporation Assay

    Spinal NG2-positive and Iba1-positive cells proliferate after PNS injury. (A) Experimental schedule and schematic diagram. Bromodeoxyuridine (50 mg/kg/day, i.p.) or vehicle was injected on days 0, 2, 4, and 6 after unilateral SNC and detected in the ipsilateral

    Journal: Journal of neuropathology and experimental neurology

    Article Title: Spinal Glia Division Contributes to Conditioning Lesion–Induced Axon Regeneration Into the Injured Spinal Cord: Potential Role of Cyclic AMP–Induced Tissue Inhibitor of Metalloproteinase-1

    doi: 10.1097/NEN.0000000000000192

    Figure Lengend Snippet: Spinal NG2-positive and Iba1-positive cells proliferate after PNS injury. (A) Experimental schedule and schematic diagram. Bromodeoxyuridine (50 mg/kg/day, i.p.) or vehicle was injected on days 0, 2, 4, and 6 after unilateral SNC and detected in the ipsilateral

    Article Snippet: The antibodies used for immunodetection were as follows: monoclonal mouse anti-BrdU (B2531; Sigma) and polyclonal rabbit anti–glial fibrillary acidic protein (GFAP; Z0334; Dako, Carpinteria, CA; for dual labeling with NG2, Iba1, and GFAP); monoclonal rat anti-BrdU (ab6326; Abcam, Cambridge, MA; for dual labeling with O1); polyclonal goat anti-CTB (703; List Biological Laboratories); polyclonal rabbit anti-NG2 (AB5320; EMD Millipore, Billerica, MA); monoclonal mouse anti-O1 (MAB344; Chemicon, Temecula, CA); monoclonal mouse anti-CD11b (MCA618R; Serotec, Raleigh, NC); polyclonal rabbit anti-Iba1 (019-19741; Wako, Richmond, VA); monoclonal mouse rat anti-CD68 (MCA341R; Serotec); polyclonal goat anti–calcitonin gene–related peptide (CGRP; 1720-9007; Biogenesis, Bournemouth, United Kingdom); and polyclonal rabbit anti-S100 (Z0311; Dako).

    Techniques: Injection

    Deletion of Misu increases dormancy of bulge stem cell. (A,B) Whole mount staining of wild-type and Misu −/− tail skin for LRC (green), keratin 14 (K14) (red) and DAPI (blue) after 4 months chase period. (C) Automated quantification of LRC in tail outer hair follicles in (A,B). (D) Frequency distributions of the intensity of the BrdU-label in LRC in (A,B). Error bars represent SEM (n = 10) from at least 3 independent experiments. (E,F) Flow cytometry for Itgα6 and CD34 in epidermis in telogen (P49) (E) and catagen (P40) (F). The percentage of double positive cells in each group is shown ± SEM (n = 3). The red lines in (E) indicate a cell population that increases in Misu −/− skin in telogen. (G,H) Epidermal cells sorted for CD34 +ve and low (L) and high (H) levels of Itgα6 were subjected to QPCR for bulge and hair germ markers as indicated (G). RNA levels were measured relative to GAPDH and presented as fold enrichment in Misu −/− mice versus wild-type controls. Error bars represent SEM (n = 3). (I) BrdU labelling and chasing regime to measure migration of LCR from bulge to hair germ at telogen at P47 (arrow). (J,K) Detection of LRC (green), Ki67 (red) and DAPI (blue) in high and low part of the bulge (straight line) and hair germ (HG) in hair follicles, indicated by dotted line, from wild-type (wt) (J) and Misu −/− mice (K). (L) Automated quantification of LRC in the whole hair follicle (total), high and low bulge region and the hair germ. Error bars indicate SEM (n = 5). (M) Percentages of hair follicles without LRC in the lower bulge region. Scale bars: 250 µm (A,B); 50 µm (J,K).

    Journal: PLoS Genetics

    Article Title: The RNA-Methyltransferase Misu (NSun2) Poises Epidermal Stem Cells to Differentiate

    doi: 10.1371/journal.pgen.1002403

    Figure Lengend Snippet: Deletion of Misu increases dormancy of bulge stem cell. (A,B) Whole mount staining of wild-type and Misu −/− tail skin for LRC (green), keratin 14 (K14) (red) and DAPI (blue) after 4 months chase period. (C) Automated quantification of LRC in tail outer hair follicles in (A,B). (D) Frequency distributions of the intensity of the BrdU-label in LRC in (A,B). Error bars represent SEM (n = 10) from at least 3 independent experiments. (E,F) Flow cytometry for Itgα6 and CD34 in epidermis in telogen (P49) (E) and catagen (P40) (F). The percentage of double positive cells in each group is shown ± SEM (n = 3). The red lines in (E) indicate a cell population that increases in Misu −/− skin in telogen. (G,H) Epidermal cells sorted for CD34 +ve and low (L) and high (H) levels of Itgα6 were subjected to QPCR for bulge and hair germ markers as indicated (G). RNA levels were measured relative to GAPDH and presented as fold enrichment in Misu −/− mice versus wild-type controls. Error bars represent SEM (n = 3). (I) BrdU labelling and chasing regime to measure migration of LCR from bulge to hair germ at telogen at P47 (arrow). (J,K) Detection of LRC (green), Ki67 (red) and DAPI (blue) in high and low part of the bulge (straight line) and hair germ (HG) in hair follicles, indicated by dotted line, from wild-type (wt) (J) and Misu −/− mice (K). (L) Automated quantification of LRC in the whole hair follicle (total), high and low bulge region and the hair germ. Error bars indicate SEM (n = 5). (M) Percentages of hair follicles without LRC in the lower bulge region. Scale bars: 250 µm (A,B); 50 µm (J,K).

    Article Snippet: Primary antibodies were used at the following dilutions: rabbit monoclonal antibody to Ki67 (1∶100; SP6, Vector Labs), rabbit polyclonal anti mouse keratin 14 (1∶2000; Covance), rabbit polyclonal anti mouse keratin 10 (1∶500; Covance), mouse monoclonal anti Gata3 (1∶50; HG3-31, Santa Cruz Biotechnology), rabbit polyclonal anti Lef1 (1∶50; Cell Signaling Technology), mouse polyclonal anti Dlx3 (1∶200; Abnova), guinea pig polyclonal anti hair keratins 31, 71 and 72 (1∶200; Progen), rabbit polyclonal anti keratin 6 (1∶5.000; Babco), rabbit polyclonal to phosphor-Smad1/5/8 (1∶50; Cell Signaling Technology), rabbit polyclonal CUK-1079-A antibody to mouse Misu (1∶1000; produced by Covalab), mouse monoclonal to keratin 15 (1∶1000) ), rat monoclonal anti BrdU (1∶100; Abcam), goat polyclonal anti P-cadherin (1∶100; R & D Systems), rat monoclonal anti α6 integrin (1∶500; GoH3, AbD Serotec), Secondary antibodies (Alexa Fluor 594- and 488-conjugated anti-rabbit, mouse, rat and guinea pig, Invitrogen) were added at a dilution of 1∶500 for 1 hour at room temperature together with DAPI to label nuclei.

    Techniques: Staining, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, Mouse Assay, Migration

    Activity-induced changes in the number of BrdU-labeled cells and new neurons in the dentate gyrus . (A,B) Confocal projections of the dentate gyrus (z-stack of 10 optical sections with 1.7-μm thickness). BrdU, Red; DCX, Blue. Scale bar, 25 μm in (A–D) Confocal image (optical section of 1-μm thickness) of STDSTD in (C) and RUNENR in (D) showing NeuN, Green; BrdU, Red, S100β, Blue; Inset: a, NeuN; b, BrdU; c, S100β. (D) Arrowheads in red indicating colocalization of BrdU and NeuN. Scale bar, 25 μm in (C,D) . (E–H) Brightfield images of BrdU-positive cells in the dentate gyrus. Scale bar (in E for E–H ), 100 μm.

    Journal: Frontiers in Neuroscience

    Article Title: Additive Effects of Physical Exercise and Environmental Enrichment on Adult Hippocampal Neurogenesis in Mice

    doi: 10.3389/neuro.22.002.2009

    Figure Lengend Snippet: Activity-induced changes in the number of BrdU-labeled cells and new neurons in the dentate gyrus . (A,B) Confocal projections of the dentate gyrus (z-stack of 10 optical sections with 1.7-μm thickness). BrdU, Red; DCX, Blue. Scale bar, 25 μm in (A–D) Confocal image (optical section of 1-μm thickness) of STDSTD in (C) and RUNENR in (D) showing NeuN, Green; BrdU, Red, S100β, Blue; Inset: a, NeuN; b, BrdU; c, S100β. (D) Arrowheads in red indicating colocalization of BrdU and NeuN. Scale bar, 25 μm in (C,D) . (E–H) Brightfield images of BrdU-positive cells in the dentate gyrus. Scale bar (in E for E–H ), 100 μm.

    Article Snippet: We here used the following primary antibodies: rat anti-BrdU (Harlan Seralab) 1:500, mouse anti-NeuN (Chemicon) 1:100, goat anti-Dcx (Santa Cruz) 1: 250, rabbit anti-S100β (Swant) 1:2000.

    Techniques: Activity Assay, Labeling

    Haloperidol increases the number of CP cells in the subependyma, neurogenesis in the olfactory bulbs, and gliogenesis in the striatum. A , Left, Photomicrograph of progenitor cells after haloperidol treatment. Right, Haloperidol increased BrdU (+) cell numbers in lateral ventricle subependyma. B , Left, Photomicrograph of neurogenesis in the olfactory bulb. Right, Haloperidol increased BrdU (+) /NeuN (+) cell numbers in the olfactory bulbs. C , Left, Photomicrograph of non-neuronal cell genesis in the striatum. Right, Haloperidol increased BrdU (+) /NeuN (-) cell numbers in the striatum. Single and double arrowheads indicate BrdU (+) /NeuN (-) and BrdU (+) /NeuN (+) cells, respectively. All data are presented as mean ± SEM. Scale bars, 50 μm. * p

    Journal: The Journal of Neuroscience

    Article Title: Dopamine Specifically Inhibits Forebrain Neural Stem Cell Proliferation, Suggesting a Novel Effect of Antipsychotic Drugs

    doi: 10.1523/JNEUROSCI.1120-05.2005

    Figure Lengend Snippet: Haloperidol increases the number of CP cells in the subependyma, neurogenesis in the olfactory bulbs, and gliogenesis in the striatum. A , Left, Photomicrograph of progenitor cells after haloperidol treatment. Right, Haloperidol increased BrdU (+) cell numbers in lateral ventricle subependyma. B , Left, Photomicrograph of neurogenesis in the olfactory bulb. Right, Haloperidol increased BrdU (+) /NeuN (+) cell numbers in the olfactory bulbs. C , Left, Photomicrograph of non-neuronal cell genesis in the striatum. Right, Haloperidol increased BrdU (+) /NeuN (-) cell numbers in the striatum. Single and double arrowheads indicate BrdU (+) /NeuN (-) and BrdU (+) /NeuN (+) cells, respectively. All data are presented as mean ± SEM. Scale bars, 50 μm. * p

    Article Snippet: Animals were killed, tissue was prepared, and immunostaining for BrdU and neuronal-specific nuclear protein (NeuN) was performed as reported previously ( ) using the following reagents: rat anti-BrdU antibody (1:100; Harlan Sera-Lab, Loughborough, UK), FITC donkey anti-rat antibody (1:200; Jackson ImmunoResearch, West Grove, PA), mouse anti-NeuN (1:200; Chemicon, Temecula, CA), and Alexa Fluor 555 goat anti-mouse (1:300; Molecular Probes, Eugene, OR).

    Techniques:

    S-phase labeling with BrdU IHC labeling for BrdU identified newborn cells in the subcortical white matter at 18 hours (Proliferation group, A and B) and four weeks (Survival group, C and D) after BrdU injection in GH/IGF-I replete (A and C) and deficient (B and D) rats. The cingulate cortex (Cg) and hippocampus (Hp) are identified in each section for orientation. Inset: medial CC at higher magnification. Scale bar = 250 µm, 50 µm (inset).

    Journal: Glia

    Article Title: Adult-Onset Deficiency in Growth Hormone and Insulin-Like Growth Factor-I Alters Oligodendrocyte Turnover in the Corpus Callosum

    doi: 10.1002/glia.20829

    Figure Lengend Snippet: S-phase labeling with BrdU IHC labeling for BrdU identified newborn cells in the subcortical white matter at 18 hours (Proliferation group, A and B) and four weeks (Survival group, C and D) after BrdU injection in GH/IGF-I replete (A and C) and deficient (B and D) rats. The cingulate cortex (Cg) and hippocampus (Hp) are identified in each section for orientation. Inset: medial CC at higher magnification. Scale bar = 250 µm, 50 µm (inset).

    Article Snippet: The density of BrdU-labeled cells was analyzed in sections labeled by IHC using rat monoclonal anti-BrdU (2.5 µg/ml; Accurate Chemical & Scientific Corp., see ).

    Techniques: Labeling, Immunohistochemistry, Injection

    Pulse-chase analysis of basal cell proliferation. A–C: Dual immunofluorescence analysis of BrdU and keratin 5 in naphthalene-injured tracheas that were pulsed with BrdU on recovery day three and killed three hours later (day three) or after a three- (day six) or a six-day chase (day nine). Yellow arrows indicate BrdU+/keratin 5+ dual positive cells. White arrows indicate BrdU+ cells that are keratin 5−. Adjacent serial sections to those presented in A–C are labeled d3, d6, and d9, respectively. These sections were stained for keratin 14, CCSP, or ACT. Examples of antigen-positive cells are indicated by red arrows . Scale bar in A = 50 μm.

    Journal: The American Journal of Pathology

    Article Title: Tracheal Basal Cells

    doi: 10.2353/ajpath.2010.090870

    Figure Lengend Snippet: Pulse-chase analysis of basal cell proliferation. A–C: Dual immunofluorescence analysis of BrdU and keratin 5 in naphthalene-injured tracheas that were pulsed with BrdU on recovery day three and killed three hours later (day three) or after a three- (day six) or a six-day chase (day nine). Yellow arrows indicate BrdU+/keratin 5+ dual positive cells. White arrows indicate BrdU+ cells that are keratin 5−. Adjacent serial sections to those presented in A–C are labeled d3, d6, and d9, respectively. These sections were stained for keratin 14, CCSP, or ACT. Examples of antigen-positive cells are indicated by red arrows . Scale bar in A = 50 μm.

    Article Snippet: Antigens were detected with: Rabbit anti-Keratin 5 (Covance, Princeton, NJ; 1:1000), Mouse IgG3 anti-Keratin 14 (ThermoFisher, Waltham MA; 1:500), Chicken anti-Keratin 15 (Covance, 1:1000), Rat anti-Ki-67 (Dako, Glostrup, Denmark; 1:500), Goat anti-CCSP, Mouse IgG2b anti-acetylated tubulin (Sigma, 1:8000), and monoclonal Rat anti-BrdU (Accurate Chemical, Westbury, NY; 1:500).

    Techniques: Pulse Chase, Immunofluorescence, Labeling, Staining, Activated Clotting Time Assay

    X-gal-negative Newly Formed Cardiomyocytes Originate from LRCs, Which Consisted of CSCs or CPCs. (A) A CPC co-expressing Sca-1 and GATA4. In the two left panels, the arrowhead indicates a Sca-1 (green) and GATA4 (red) double-positive CPC surrounded by a basement membrane of cardiomyocytes (laminin in blue). The GATA4 signal co-localized with DAPI nuclear staining (blue) in the two right panels. Arrows indicate Sca-1-positive capillaries. Scale bar, 10 μm. (B) Partial overlap of LRCs and CPCs. The hearts of 10-week-old mice that were administered BrdU during the fetal period were immunohistochemically examined. Some anti-sarcomeric α-actinin (SA-actinin)-negative and BrdU-positive LRCs were Nkx2.5-positive (arrowhead, upper panels) or pGATA4-positive (arrowhead, lower panels). Some Nkx2.5-positive (arrow, upper panels) or pGATA4-positive (arrow, lower panels) cardiomyocytes retained BrdU. Nuclei were stained with DAPI. Scale bar, 10 μm. (C) Partial overlap of LRCs and CSCs. Cardiac side population cells (CSPs) were isolated from the hearts of 10-week-old mice that had been administered BrdU during the fetal period. BrdU-positive and multi-drug-resistant protein 1(MDR1)-positive CSPs were identified (arrowheads). Scale bar, 10 μm. (D) X-gal-negative newly formed cardiomyocytes originate from LRCs. A white arrowhead indicates SA-actinin-positive and X-gal-negative LRC-derived newly formed cardiomyocytes. An arrow indicates SA-actinin- and X-gal-positive pre-existing cardiomyocytes. Scale bar, 10 μm. Note that the nuclei of both cell types retained BrdU because of their quiescence after birth. Two yellow arrowheads indicate SA-actinin-positive and X-gal-negative cardiomyocytes, the ancestral CSCs or CPCs of which circumvented the BrdU labeling because of their quiescence. (E) Cre-mediated recombination does not occur in CSPs after tamoxifen treatment. X-gal staining of the cardiomyocyte fraction (right) and CSP cell fraction (left) from a tamoxifen-treated mouse. Arrowheads indicate X-gal-negative CSPs. An arrow indicates X-gal-positive cardiomyocytes. β-galactosidase mRNA expression in cardiomyocyte suspension and sorted CSP fraction from tamoxifen-treated and -untreated mice. Scale bar, 10 μm.

    Journal: PLoS ONE

    Article Title: Leukemia Inhibitory Factor Enhances Endogenous Cardiomyocyte Regeneration after Myocardial Infarction

    doi: 10.1371/journal.pone.0156562

    Figure Lengend Snippet: X-gal-negative Newly Formed Cardiomyocytes Originate from LRCs, Which Consisted of CSCs or CPCs. (A) A CPC co-expressing Sca-1 and GATA4. In the two left panels, the arrowhead indicates a Sca-1 (green) and GATA4 (red) double-positive CPC surrounded by a basement membrane of cardiomyocytes (laminin in blue). The GATA4 signal co-localized with DAPI nuclear staining (blue) in the two right panels. Arrows indicate Sca-1-positive capillaries. Scale bar, 10 μm. (B) Partial overlap of LRCs and CPCs. The hearts of 10-week-old mice that were administered BrdU during the fetal period were immunohistochemically examined. Some anti-sarcomeric α-actinin (SA-actinin)-negative and BrdU-positive LRCs were Nkx2.5-positive (arrowhead, upper panels) or pGATA4-positive (arrowhead, lower panels). Some Nkx2.5-positive (arrow, upper panels) or pGATA4-positive (arrow, lower panels) cardiomyocytes retained BrdU. Nuclei were stained with DAPI. Scale bar, 10 μm. (C) Partial overlap of LRCs and CSCs. Cardiac side population cells (CSPs) were isolated from the hearts of 10-week-old mice that had been administered BrdU during the fetal period. BrdU-positive and multi-drug-resistant protein 1(MDR1)-positive CSPs were identified (arrowheads). Scale bar, 10 μm. (D) X-gal-negative newly formed cardiomyocytes originate from LRCs. A white arrowhead indicates SA-actinin-positive and X-gal-negative LRC-derived newly formed cardiomyocytes. An arrow indicates SA-actinin- and X-gal-positive pre-existing cardiomyocytes. Scale bar, 10 μm. Note that the nuclei of both cell types retained BrdU because of their quiescence after birth. Two yellow arrowheads indicate SA-actinin-positive and X-gal-negative cardiomyocytes, the ancestral CSCs or CPCs of which circumvented the BrdU labeling because of their quiescence. (E) Cre-mediated recombination does not occur in CSPs after tamoxifen treatment. X-gal staining of the cardiomyocyte fraction (right) and CSP cell fraction (left) from a tamoxifen-treated mouse. Arrowheads indicate X-gal-negative CSPs. An arrow indicates X-gal-positive cardiomyocytes. β-galactosidase mRNA expression in cardiomyocyte suspension and sorted CSP fraction from tamoxifen-treated and -untreated mice. Scale bar, 10 μm.

    Article Snippet: Antibodies The primary antibodies included mouse monoclonal anti-SA-actinin (1:200, A7811, Sigma–Aldrich), mouse BA-G5 monoclonal antibody specific for cardiac α-myosin heavy chain (1:100, ab50967, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-laminin (1:200, L9393, Sigma–Aldrich), mouse monoclonal anti-beta-galactosidase (1:100, 0863363, MP Biomedicals, Santa Ana, CA, USA; LLC-Cappel), rat monoclonal anti-laminin-α-2 (1:200, sc-59854, Santa Cruz Biotechnology, Dallas, TX, USA), rat monoclonal anti-BrdU (1:250, OBT0030, AbD Serotec, Raleigh, NC, USA), mouse monoclonal anti-multi-drug-resistant protein 1 (MDR1) (1:50, ALX801-002-C100, Alexis, Lausen, Switzerland), rabbit polyclonal anti-Nkx2.5 (1:500, ab35842, Abcam), rabbit polyclonal anti-phosphorylated GATA4 (anti-pGATA4) (1:500, phosho-S105, ab92585, Abcam), goat polyclonal anti-GATA4 (1:50, sc-1237, Santa Cruz Biotechnology), mouse monoclonal anti-smooth muscle actin (1:100, IS61130, DAKO, Glostrup, Denmark), rabbit polyclonal anti-phosphorylated STAT3 (p-STAT3) (1:100, #9131, Cell Signaling Technologies, Danvers, MA, USA), rabbit polyclonal phospho-p44/42 MAPK (p-ERK) (1:50, #9101, Cell Signaling Technologies, Danvers, MA, USA), rabbit polyclonal Akt1/2/3 (Ser 473)-R (p-Akt) (1:50, sc-7985-R, Santa Cruz Biotechnology), rabbit polyclonal anti-vimentin (1:20, GP59, Progen, Heidelberg, Germany), rat monoclonal anti-CD31 (1:100, 14-0311-81, eBioscience, San Diego, CA, USA), rat monoclonal anti-mouse Ly-6A/E (Sca-1) (1:100, 14-5981-81, eBioscience), rabbit polyclonal connexin 40 (1:100, AB1726, Merck Millipore, Darmstadt, Germany), and rabbit polyclonal Ki67 antibody (1:100, ab15580, Abcam).

    Techniques: Expressing, Staining, Mouse Assay, Isolation, Derivative Assay, Labeling

    LIF Increases the Number of X-gal-negative Newly Formed Cardiomyocytes and the Frequency of BrdU Incorporation. (A) Representative images of X-gal-negative cardiomyocytes in PBS- and LIF-treated CreLacZ mice. X-gal staining (left) and immunofluorescence images (right; SA-actinin, green; laminin, red; nuclei were stained with DAPI, blue) are shown. Arrows indicate X-gal-negative cardiomyocytes. Scale bar, 20 μm. (B) Number of X-gal-negative cardiomyocytes in the MI remote area (closed bar) and the MI area (open bar) in the PBS- and LIF-treated mice after MI. Asterisks indicate significant differences between two groups. *p

    Journal: PLoS ONE

    Article Title: Leukemia Inhibitory Factor Enhances Endogenous Cardiomyocyte Regeneration after Myocardial Infarction

    doi: 10.1371/journal.pone.0156562

    Figure Lengend Snippet: LIF Increases the Number of X-gal-negative Newly Formed Cardiomyocytes and the Frequency of BrdU Incorporation. (A) Representative images of X-gal-negative cardiomyocytes in PBS- and LIF-treated CreLacZ mice. X-gal staining (left) and immunofluorescence images (right; SA-actinin, green; laminin, red; nuclei were stained with DAPI, blue) are shown. Arrows indicate X-gal-negative cardiomyocytes. Scale bar, 20 μm. (B) Number of X-gal-negative cardiomyocytes in the MI remote area (closed bar) and the MI area (open bar) in the PBS- and LIF-treated mice after MI. Asterisks indicate significant differences between two groups. *p

    Article Snippet: Antibodies The primary antibodies included mouse monoclonal anti-SA-actinin (1:200, A7811, Sigma–Aldrich), mouse BA-G5 monoclonal antibody specific for cardiac α-myosin heavy chain (1:100, ab50967, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-laminin (1:200, L9393, Sigma–Aldrich), mouse monoclonal anti-beta-galactosidase (1:100, 0863363, MP Biomedicals, Santa Ana, CA, USA; LLC-Cappel), rat monoclonal anti-laminin-α-2 (1:200, sc-59854, Santa Cruz Biotechnology, Dallas, TX, USA), rat monoclonal anti-BrdU (1:250, OBT0030, AbD Serotec, Raleigh, NC, USA), mouse monoclonal anti-multi-drug-resistant protein 1 (MDR1) (1:50, ALX801-002-C100, Alexis, Lausen, Switzerland), rabbit polyclonal anti-Nkx2.5 (1:500, ab35842, Abcam), rabbit polyclonal anti-phosphorylated GATA4 (anti-pGATA4) (1:500, phosho-S105, ab92585, Abcam), goat polyclonal anti-GATA4 (1:50, sc-1237, Santa Cruz Biotechnology), mouse monoclonal anti-smooth muscle actin (1:100, IS61130, DAKO, Glostrup, Denmark), rabbit polyclonal anti-phosphorylated STAT3 (p-STAT3) (1:100, #9131, Cell Signaling Technologies, Danvers, MA, USA), rabbit polyclonal phospho-p44/42 MAPK (p-ERK) (1:50, #9101, Cell Signaling Technologies, Danvers, MA, USA), rabbit polyclonal Akt1/2/3 (Ser 473)-R (p-Akt) (1:50, sc-7985-R, Santa Cruz Biotechnology), rabbit polyclonal anti-vimentin (1:20, GP59, Progen, Heidelberg, Germany), rat monoclonal anti-CD31 (1:100, 14-0311-81, eBioscience, San Diego, CA, USA), rat monoclonal anti-mouse Ly-6A/E (Sca-1) (1:100, 14-5981-81, eBioscience), rabbit polyclonal connexin 40 (1:100, AB1726, Merck Millipore, Darmstadt, Germany), and rabbit polyclonal Ki67 antibody (1:100, ab15580, Abcam).

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

    Environmental enrichment leads to the development of individual levels of adult hippocampal neurogenesis. ( A, B ) Representative images of Ki67 immunostaining, which marks proliferating cells in control (CTRL) and enriched (ENR) mice. ( C, D ) New-born cells were identified by BrdU immunoreactivity three weeks after the injection of BrdU. ( E ) The proportions of new-born neurons and astrocytes were determined by co-localization of BrdU (green) with NeuN (blue) and S100β (red), respectively. The image shows a single optical section. The arrowhead highlights a new-born neuron. ( F ) No difference in the number of proliferating cells can be observed between mice housed under CTRL and ENR conditions. ( G–I ) ENR mice have significantly higher means and variances in the numbers of new-born BrdU-positive cells ( G ) and new neurons ( H ), whereas only the variance of the number of new astrocytes was increased ( I ). Scale bars are as follows: ( A–D ) 100 µm; (E), 50 µm; (E inset), 10 µm. Box and whisker plots, see Figure 2 . Asterisks indicate significant effects at a 5% threshold.

    Journal: eLife

    Article Title: Selective increases in inter-individual variability in response to environmental enrichment in female mice

    doi: 10.7554/eLife.35690

    Figure Lengend Snippet: Environmental enrichment leads to the development of individual levels of adult hippocampal neurogenesis. ( A, B ) Representative images of Ki67 immunostaining, which marks proliferating cells in control (CTRL) and enriched (ENR) mice. ( C, D ) New-born cells were identified by BrdU immunoreactivity three weeks after the injection of BrdU. ( E ) The proportions of new-born neurons and astrocytes were determined by co-localization of BrdU (green) with NeuN (blue) and S100β (red), respectively. The image shows a single optical section. The arrowhead highlights a new-born neuron. ( F ) No difference in the number of proliferating cells can be observed between mice housed under CTRL and ENR conditions. ( G–I ) ENR mice have significantly higher means and variances in the numbers of new-born BrdU-positive cells ( G ) and new neurons ( H ), whereas only the variance of the number of new astrocytes was increased ( I ). Scale bars are as follows: ( A–D ) 100 µm; (E), 50 µm; (E inset), 10 µm. Box and whisker plots, see Figure 2 . Asterisks indicate significant effects at a 5% threshold.

    Article Snippet: Primary antibodies were applied overnight at 4°C as follows: monoclonal rat anti-BrdU (1:500, Serotec), rabbit anti-Ki67 (Novocastra, 1:500), and rabbit anti-Synaptoporin (Synaptic Systems, 1:500).

    Techniques: Immunostaining, Mouse Assay, Injection, Whisker Assay

    VIP enhances colonic crypt function at baseline. Exogenous VIP was administered to VIPKO mice (VIPKO-VIP) daily for 10 days. Immunostaining for Ki67 and quantitative analysis of Ki67+ve cells (A). Immunostaining for BrdU+ve cells and calculated spatial distribution at 72h post injection (B). Quantitative analysis of TUNEL +ve crypt IEC (C). Epithelial permeability measured by FITC dextran (D); n = 6–10 animals/group, results are represented as means ± SEM, *P

    Journal: PLoS ONE

    Article Title: Vasoactive Intestinal Polypeptide Promotes Intestinal Barrier Homeostasis and Protection Against Colitis in Mice

    doi: 10.1371/journal.pone.0125225

    Figure Lengend Snippet: VIP enhances colonic crypt function at baseline. Exogenous VIP was administered to VIPKO mice (VIPKO-VIP) daily for 10 days. Immunostaining for Ki67 and quantitative analysis of Ki67+ve cells (A). Immunostaining for BrdU+ve cells and calculated spatial distribution at 72h post injection (B). Quantitative analysis of TUNEL +ve crypt IEC (C). Epithelial permeability measured by FITC dextran (D); n = 6–10 animals/group, results are represented as means ± SEM, *P

    Article Snippet: Primary antibodies included rabbit monoclonal anti-ki67 (Thermo scientific), rabbit polyclonal antisera against murine colonic mucin Muc2 (1:50; a gift from Jan Dekker), rabbit polyclonal antisera against TFF3 (1:200; a gift from D. Podolsky), polyclonal rabbit anti-VIP (Immunostar), goat polyclonal anti-carbonic anhydrase (CA) I (Santa Cruz), rat anti-BrdU monoclonal antibody (1:200, AbD Serotec) and rabbit anti-serotonin (5HT) (Antibodies incorporated).

    Techniques: Mouse Assay, Immunostaining, Injection, TUNEL Assay, Permeability

    Fluorescence microscopy of neuronal and microglial cells derived from dividing progenitors. Panel A is a confocal photomicrograph that shows NeuN+ (red) cells in the cortex of a control mouse brain for the striatal line. Panel B is a confocal photomicrograph that shows a BrdU+ (green) cell in the same field. Panel C represents the merging of panels A and B , and indicates that the cell (arrow) is NeuN+/BrdU+ (yellow) and therefore represents a neuron derived from a dividing progenitor cell. Scale bar in panels A , B and C represents 12 μm. Panel D is a confocal photomicrograph that shows Iba1+ (red) cells in the striatum of a mutant mouse brain from the striatal line. Panel E is a confocal photomicrograph that shows BrdU+ (green) cells in the same field. Panel F represents the merging of panels D and E and indicates that one cell (arrow) is Iba1+/BrdU+ (yellow) and therefore represents a microglial cell derived from a cell that has undergone cell division. Scale bar in panels D , E and F represents 15 μm.

    Journal: BMC Neuroscience

    Article Title: Phenotyping dividing cells in mouse models of neurodegenerative basal ganglia diseases

    doi: 10.1186/1471-2202-14-111

    Figure Lengend Snippet: Fluorescence microscopy of neuronal and microglial cells derived from dividing progenitors. Panel A is a confocal photomicrograph that shows NeuN+ (red) cells in the cortex of a control mouse brain for the striatal line. Panel B is a confocal photomicrograph that shows a BrdU+ (green) cell in the same field. Panel C represents the merging of panels A and B , and indicates that the cell (arrow) is NeuN+/BrdU+ (yellow) and therefore represents a neuron derived from a dividing progenitor cell. Scale bar in panels A , B and C represents 12 μm. Panel D is a confocal photomicrograph that shows Iba1+ (red) cells in the striatum of a mutant mouse brain from the striatal line. Panel E is a confocal photomicrograph that shows BrdU+ (green) cells in the same field. Panel F represents the merging of panels D and E and indicates that one cell (arrow) is Iba1+/BrdU+ (yellow) and therefore represents a microglial cell derived from a cell that has undergone cell division. Scale bar in panels D , E and F represents 15 μm.

    Article Snippet: Double labelling using rat anti-BrdU and mouse anti-NeuN The staining protocol required the following modification in the case of double labelling using rat anti-BrdU and mouse anti-NeuN.

    Techniques: Fluorescence, Microscopy, Derivative Assay, Mutagenesis

    Regional quantification of neuronal, microglial, astroglial and oligodendroglial lineage cells derived from dividing progenitors in GFP-control and GFP-global mutant mice. Graphs showing the number of double-positive NeuN+/BrdU+, Iba1+/BrdU+, S100β+/BrdU+ and Olig2+/BrdU+ cells at specific sites relative to bregma in the M1 motor cortex (A-D) , striatum (E-H) and periventricular (I-L) regions of the brain of mice belonging to the GFP-global line and GFP-control mice injected for 2 weeks with BrdU+ at 4 weeks of age and killed at age 8 weeks of age for analysis. Each point represents the mean ± SEM as determined for each bregma level in mutant (n=4) and control (n=4) mice. There was a greater number of NeuN+/BrdU+ cells in GFP-global mutant relative to control mice measured at 0.26mm from bregma (** P

    Journal: BMC Neuroscience

    Article Title: Phenotyping dividing cells in mouse models of neurodegenerative basal ganglia diseases

    doi: 10.1186/1471-2202-14-111

    Figure Lengend Snippet: Regional quantification of neuronal, microglial, astroglial and oligodendroglial lineage cells derived from dividing progenitors in GFP-control and GFP-global mutant mice. Graphs showing the number of double-positive NeuN+/BrdU+, Iba1+/BrdU+, S100β+/BrdU+ and Olig2+/BrdU+ cells at specific sites relative to bregma in the M1 motor cortex (A-D) , striatum (E-H) and periventricular (I-L) regions of the brain of mice belonging to the GFP-global line and GFP-control mice injected for 2 weeks with BrdU+ at 4 weeks of age and killed at age 8 weeks of age for analysis. Each point represents the mean ± SEM as determined for each bregma level in mutant (n=4) and control (n=4) mice. There was a greater number of NeuN+/BrdU+ cells in GFP-global mutant relative to control mice measured at 0.26mm from bregma (** P

    Article Snippet: Double labelling using rat anti-BrdU and mouse anti-NeuN The staining protocol required the following modification in the case of double labelling using rat anti-BrdU and mouse anti-NeuN.

    Techniques: Derivative Assay, Mutagenesis, Mouse Assay, Injection

    Regional quantification of neuronal, microglial, astroglial, and oligodendroglial cells derived from dividing progenitors in control and striatal line mutant mice. Graphs showing the number of double-positive NeuN+/BrdU+, Iba1+/BrdU+, S100β+/BrdU+ and Olig2+/BrdU+ cells at specific sites relative to bregma in the M1 motor cortex (A-D) , striatum (E-H) and periventricular (I-L) regions of the brain of mice belonging to the striatal line and control mice injected for 2 weeks with BrdU+ at 4 weeks of age and killed at age 8 weeks. There was a significant increase in the number of Iba1+/BrdU+ cells in the striatum and periventricular regions (both P

    Journal: BMC Neuroscience

    Article Title: Phenotyping dividing cells in mouse models of neurodegenerative basal ganglia diseases

    doi: 10.1186/1471-2202-14-111

    Figure Lengend Snippet: Regional quantification of neuronal, microglial, astroglial, and oligodendroglial cells derived from dividing progenitors in control and striatal line mutant mice. Graphs showing the number of double-positive NeuN+/BrdU+, Iba1+/BrdU+, S100β+/BrdU+ and Olig2+/BrdU+ cells at specific sites relative to bregma in the M1 motor cortex (A-D) , striatum (E-H) and periventricular (I-L) regions of the brain of mice belonging to the striatal line and control mice injected for 2 weeks with BrdU+ at 4 weeks of age and killed at age 8 weeks. There was a significant increase in the number of Iba1+/BrdU+ cells in the striatum and periventricular regions (both P

    Article Snippet: Double labelling using rat anti-BrdU and mouse anti-NeuN The staining protocol required the following modification in the case of double labelling using rat anti-BrdU and mouse anti-NeuN.

    Techniques: Derivative Assay, Mutagenesis, Mouse Assay, Injection

    Depletion of slowly dividing neural stem cells and impairment of regeneration after cytosine-β-D-arabinofuranoside (AraC) treatment in Hes1/3/5/Hey1 mutant adult mouse brains. ( A ) Experimental design. ( B – E ) Dividing progenitors ( t = 0; B , C ) and slowly dividing cells, ones that retained labeling even after 12 d ( t = 12; D , E ), in the SVZ were labeled with BrdU in control ( B , D ) and Hes1/3/5/Hey1 mutant ( C , E ) mice. ( F , G ) Quantification of the number of BrdU-incorporating cells in the SVZ at t = 0 and t = 12 after BrdU administration for 14 consecutive days. WT-control ( Hes1 +/+ ;Hes3 +/+ ;Hes5 +/+ ;Hey1 +/+ ) mice are shown as a reference. ( H ) Experimental design. ( I – P ) Coronal sections of the SVZ showing BrdU-incorporating cells soon after the removal of the osmotic pump ( t = 0; I , J , M , N ) and 10 d after the pump removal ( t = 10; K , L , O , P ) in AraC-treated control ( I – L ) and AraC-treated Hes1/3/5/Hey1 mutant ( M – P ) mice. ( Q ) Quantification of the number of BrdU-incorporating cells in the SVZ. BrdU was administered for 4 h before sacrifice. (*) P

    Journal: Genes & Development

    Article Title: High Hes1 expression and resultant Ascl1 suppression regulate quiescent vs. active neural stem cells in the adult mouse brain

    doi: 10.1101/gad.323196.118

    Figure Lengend Snippet: Depletion of slowly dividing neural stem cells and impairment of regeneration after cytosine-β-D-arabinofuranoside (AraC) treatment in Hes1/3/5/Hey1 mutant adult mouse brains. ( A ) Experimental design. ( B – E ) Dividing progenitors ( t = 0; B , C ) and slowly dividing cells, ones that retained labeling even after 12 d ( t = 12; D , E ), in the SVZ were labeled with BrdU in control ( B , D ) and Hes1/3/5/Hey1 mutant ( C , E ) mice. ( F , G ) Quantification of the number of BrdU-incorporating cells in the SVZ at t = 0 and t = 12 after BrdU administration for 14 consecutive days. WT-control ( Hes1 +/+ ;Hes3 +/+ ;Hes5 +/+ ;Hey1 +/+ ) mice are shown as a reference. ( H ) Experimental design. ( I – P ) Coronal sections of the SVZ showing BrdU-incorporating cells soon after the removal of the osmotic pump ( t = 0; I , J , M , N ) and 10 d after the pump removal ( t = 10; K , L , O , P ) in AraC-treated control ( I – L ) and AraC-treated Hes1/3/5/Hey1 mutant ( M – P ) mice. ( Q ) Quantification of the number of BrdU-incorporating cells in the SVZ. BrdU was administered for 4 h before sacrifice. (*) P

    Article Snippet: The following primary antibodies (final dilution and source) were used: rabbit anti-Hes1 (1:500) , mouse anti-βIII-tubulin (1:500; Babco), rat anti-BrdU (1:50; Oxford Biotech), goat antidoublecortin (DCX; 1:200; Santa Cruz Biotechnology), mouse anti-GFAP (1:200; Sigma), rabbit anti-GFAP (1:200; Sigma), mouse antimammalian achaete–schute homolog 1 (1:20; BD Pharmingen), mouse anti-Nestin (1:200; BD Pharmingen), rabbit anti-MCM2 (1:500; Abcam), mouse anti-cyclinD1 (1:200; Santa Cruz Biotechnology), goat anti-Sox2 (1:500; R & D Systems), rat anti-GFP (1:500; Nacalai Tesque), chicken anti-GFP (1:500; Abcam), and mouse anti-Ki67 (1:50; BD Biosciences).

    Techniques: Mutagenesis, Labeling, Mouse Assay

    Up-regulation of Ascl1 and premature loss of neural stem cells in the adult brains of Hes1/3/5/Hey1 mutant mice. Coronal sections of the SVZ of the lateral ventricles ( A – Z ) and the SGZ of the hippocampal dentate gyrus ( AA–AO ) of Hes1 floxed/floxed ;Hes3 −/− ;Hes5 −/− ;Hey1 −/− mice (control) and Hes1/3/5/Hey1 mutant mice were examined by immunohistochemistry. BrdU was administered for 2 h before sacrifice. ( A – H ) Control ( A – D ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 10 d before ( E – H ). ( I – P ) Control ( I – L ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk before ( M – P ). ( Q – X ) Control ( Q – T ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 mo before ( U – X ). Boxed regions in R and V are enlarged at the right . ( Y , Z ) Quantification of BrdU + ( Y ) and Ascl1 + ( Z ) cells in control and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 10 d ( left ), 3 wk ( middle ), and 3 mo ( right ) before BrdU administration. ( AA–AN ) Control ( AA–AC , AG–AJ ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk ( AD–AF ) and 3 mo ( AK–AN ) before. ( AJ , AN ) BrdU was given for seven consecutive days before sacrifice. ( AO ) Quantification of DCX + , GFAP + ;MCM2 + , and BrdU + cells in control and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk ( left ) and 3 mo ( right ) before. (*) P

    Journal: Genes & Development

    Article Title: High Hes1 expression and resultant Ascl1 suppression regulate quiescent vs. active neural stem cells in the adult mouse brain

    doi: 10.1101/gad.323196.118

    Figure Lengend Snippet: Up-regulation of Ascl1 and premature loss of neural stem cells in the adult brains of Hes1/3/5/Hey1 mutant mice. Coronal sections of the SVZ of the lateral ventricles ( A – Z ) and the SGZ of the hippocampal dentate gyrus ( AA–AO ) of Hes1 floxed/floxed ;Hes3 −/− ;Hes5 −/− ;Hey1 −/− mice (control) and Hes1/3/5/Hey1 mutant mice were examined by immunohistochemistry. BrdU was administered for 2 h before sacrifice. ( A – H ) Control ( A – D ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 10 d before ( E – H ). ( I – P ) Control ( I – L ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk before ( M – P ). ( Q – X ) Control ( Q – T ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 mo before ( U – X ). Boxed regions in R and V are enlarged at the right . ( Y , Z ) Quantification of BrdU + ( Y ) and Ascl1 + ( Z ) cells in control and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 10 d ( left ), 3 wk ( middle ), and 3 mo ( right ) before BrdU administration. ( AA–AN ) Control ( AA–AC , AG–AJ ) and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk ( AD–AF ) and 3 mo ( AK–AN ) before. ( AJ , AN ) BrdU was given for seven consecutive days before sacrifice. ( AO ) Quantification of DCX + , GFAP + ;MCM2 + , and BrdU + cells in control and Hes1/3/5/Hey1 mutant mice that were treated with tamoxifen 3 wk ( left ) and 3 mo ( right ) before. (*) P

    Article Snippet: The following primary antibodies (final dilution and source) were used: rabbit anti-Hes1 (1:500) , mouse anti-βIII-tubulin (1:500; Babco), rat anti-BrdU (1:50; Oxford Biotech), goat antidoublecortin (DCX; 1:200; Santa Cruz Biotechnology), mouse anti-GFAP (1:200; Sigma), rabbit anti-GFAP (1:200; Sigma), mouse antimammalian achaete–schute homolog 1 (1:20; BD Pharmingen), mouse anti-Nestin (1:200; BD Pharmingen), rabbit anti-MCM2 (1:500; Abcam), mouse anti-cyclinD1 (1:200; Santa Cruz Biotechnology), goat anti-Sox2 (1:500; R & D Systems), rat anti-GFP (1:500; Nacalai Tesque), chicken anti-GFP (1:500; Abcam), and mouse anti-Ki67 (1:50; BD Biosciences).

    Techniques: Mutagenesis, Mouse Assay, Immunohistochemistry

    Suppression of Ascl1 expression and neurogenesis in the adult mouse brain by sustained expression of Hes1. ( A ) Experimental design. ( B – I ) Immunohistological analysis of the SVZ of R26-CFP ( B – E ) and R26-Hes1-iresGFP ( F – I ) mice that had been treated with tamoxifen 1 wk before. BrdU was administered for 2 h before sacrifice. ( J , K ) Quantification of BrdU-incorporating ( J ) and Ascl1-expressing ( K ) cells among CFP- or GFP-labeled cells. ( L ) Structures of lentiviruses. ( M – T ) Immunohistological analysis of the SGZ 1 wk after infection with EF-Venus virus ( M – P ) or EF-Hes1-Venus virus ( Q – T ). ( U ) Quantification of MCM2 + cells among virus-infected cells (Venus + ). (*) P

    Journal: Genes & Development

    Article Title: High Hes1 expression and resultant Ascl1 suppression regulate quiescent vs. active neural stem cells in the adult mouse brain

    doi: 10.1101/gad.323196.118

    Figure Lengend Snippet: Suppression of Ascl1 expression and neurogenesis in the adult mouse brain by sustained expression of Hes1. ( A ) Experimental design. ( B – I ) Immunohistological analysis of the SVZ of R26-CFP ( B – E ) and R26-Hes1-iresGFP ( F – I ) mice that had been treated with tamoxifen 1 wk before. BrdU was administered for 2 h before sacrifice. ( J , K ) Quantification of BrdU-incorporating ( J ) and Ascl1-expressing ( K ) cells among CFP- or GFP-labeled cells. ( L ) Structures of lentiviruses. ( M – T ) Immunohistological analysis of the SGZ 1 wk after infection with EF-Venus virus ( M – P ) or EF-Hes1-Venus virus ( Q – T ). ( U ) Quantification of MCM2 + cells among virus-infected cells (Venus + ). (*) P

    Article Snippet: The following primary antibodies (final dilution and source) were used: rabbit anti-Hes1 (1:500) , mouse anti-βIII-tubulin (1:500; Babco), rat anti-BrdU (1:50; Oxford Biotech), goat antidoublecortin (DCX; 1:200; Santa Cruz Biotechnology), mouse anti-GFAP (1:200; Sigma), rabbit anti-GFAP (1:200; Sigma), mouse antimammalian achaete–schute homolog 1 (1:20; BD Pharmingen), mouse anti-Nestin (1:200; BD Pharmingen), rabbit anti-MCM2 (1:500; Abcam), mouse anti-cyclinD1 (1:200; Santa Cruz Biotechnology), goat anti-Sox2 (1:500; R & D Systems), rat anti-GFP (1:500; Nacalai Tesque), chicken anti-GFP (1:500; Abcam), and mouse anti-Ki67 (1:50; BD Biosciences).

    Techniques: Expressing, Mouse Assay, Labeling, Infection

    CM-specific overexpression of dn-c-kit resulted in CM hypertrophy, leading to a thicker LV wall, a smaller LV cavity and higher ejection fraction. ( a ) Expression of Kit (c-Kit) mRNA in male P196 dn-c-kit-Tg (Tg; n = 11) and NTL (n = 6) hearts; differences analysed by Student’s t -test. (b , c ) Examples (white arrowheads) of BrdU + /cTnT + ( b ) or H3P + /cTnT + ( c ) CMs isolated from P121 Tg ( b , right panel, and c ) or NTL ( b , left panel) hearts after 9 days of BrdU (10 mg/kg/day) infusion by osmotic mini-pump. Scale bar: 50 μm. ( d ) Left panels, representative photographs, and right panel, quantitation of surface area showing hypertrophy of isolated CMs from P121 Tg (682 CMs analysed from n = 3 mice) relative to NTL hearts (1015 CMs analysed from n = 3 mice); differences analysed by Student’s t -test. Scale bar: 100 μm. ( e ) Abundance of Myh6 (encoding α-MHC) and Acta1 (α-skeletal actin) mRNAs was similar, while abundance of other mRNAs ( Nppa (ANP), Nppb (BNP), Myh 7 (β-MHC)) and the Myh 7 :Myh6 (β-:α-MHC) ratio were significantly increased in male P196 dn-c-kit-Tg relative to NTL hearts (n = 7); differences analysed by Student’s t -test. ( f ) Increased LV wall thickness to chamber radius ( h/r ) ratio at end-diastole in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test. ( g ) Increased ejection fraction in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test.

    Journal: Scientific Reports

    Article Title: Cardiac hypertrophy limits infarct expansion after myocardial infarction in mice

    doi: 10.1038/s41598-018-24525-6

    Figure Lengend Snippet: CM-specific overexpression of dn-c-kit resulted in CM hypertrophy, leading to a thicker LV wall, a smaller LV cavity and higher ejection fraction. ( a ) Expression of Kit (c-Kit) mRNA in male P196 dn-c-kit-Tg (Tg; n = 11) and NTL (n = 6) hearts; differences analysed by Student’s t -test. (b , c ) Examples (white arrowheads) of BrdU + /cTnT + ( b ) or H3P + /cTnT + ( c ) CMs isolated from P121 Tg ( b , right panel, and c ) or NTL ( b , left panel) hearts after 9 days of BrdU (10 mg/kg/day) infusion by osmotic mini-pump. Scale bar: 50 μm. ( d ) Left panels, representative photographs, and right panel, quantitation of surface area showing hypertrophy of isolated CMs from P121 Tg (682 CMs analysed from n = 3 mice) relative to NTL hearts (1015 CMs analysed from n = 3 mice); differences analysed by Student’s t -test. Scale bar: 100 μm. ( e ) Abundance of Myh6 (encoding α-MHC) and Acta1 (α-skeletal actin) mRNAs was similar, while abundance of other mRNAs ( Nppa (ANP), Nppb (BNP), Myh 7 (β-MHC)) and the Myh 7 :Myh6 (β-:α-MHC) ratio were significantly increased in male P196 dn-c-kit-Tg relative to NTL hearts (n = 7); differences analysed by Student’s t -test. ( f ) Increased LV wall thickness to chamber radius ( h/r ) ratio at end-diastole in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test. ( g ) Increased ejection fraction in P ≥ 196 dn-c-kit-Tg (n = 10–11) relative to NTL (n = 9) hearts; differences analysed by two-way ANOVA with Tukey’s multiple comparison test.

    Article Snippet: All slides were blocked and permeabilized for 1 h (for BrdU or H3P staining: 5% (v/v) normal donkey serum, 0.2% PBS-Triton X-100 (PBS-T); for AurB staining: 3% (v/v) normal donkey serum, 3% BSA (v/v), 0.1% PBS-T) before incubating overnight at 4 °C with primary mouse anti-cTnT monoclonal antibody (Abcam, ab10214, 1:600) and rat anti-BrdU monoclonal antibody (Abcam, ab 6326, 1:100), rabbit anti-H3P polyclonal antibody (Abcam, ab5176, 1:60) or rabbit anti-AurB polyclonal antibody (Abcam, ab2254, 1:50).

    Techniques: Over Expression, Expressing, Isolation, Quantitation Assay, Mouse Assay, Aqueous Normal-phase Chromatography

    p107 regulates the neural precursor population through the repression of Hes1. (a) Adult mice received intraperitoneal injections of BrdU to label proliferating cells over a 10-h period. BrdU-positive cells were counted in every 10th section through the forebrains of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 4), and p107 −/− ( n = 5) mice. (b) Proliferating progenitors in E10.5 brains were identified by PH3 immunohistochemistry. PH3-positive cells were counted in three representative sections of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 3), and p107 −/− ( n = 3) brains. (c) A 2-h BrdU pulse labeled proliferating progenitors in E13.5 brains. BrdU-positive cells were counted in four representative regions of the brain in wild-type ( n = 4), Hes1 +/− :p107 −/− ( n = 4), Hes1 +/− ( n = 4), and p107 −/− ( n = 3) embryos. Note that loss of a single Hes1 allele restored the numbers of progenitor cells to wild-type levels in p107 −/− mice at embryonic and adult ages. Means were statistically analyzed by one-way analysis of variance followed by Tukey's individual comparison of the means. Error bars represent SEM. *, P

    Journal: The Journal of Cell Biology

    Article Title: The Retinoblastoma family member p107 regulates the rate of progenitor commitment to a neuronal fate

    doi: 10.1083/jcb.200703176

    Figure Lengend Snippet: p107 regulates the neural precursor population through the repression of Hes1. (a) Adult mice received intraperitoneal injections of BrdU to label proliferating cells over a 10-h period. BrdU-positive cells were counted in every 10th section through the forebrains of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 4), and p107 −/− ( n = 5) mice. (b) Proliferating progenitors in E10.5 brains were identified by PH3 immunohistochemistry. PH3-positive cells were counted in three representative sections of wild-type ( n = 3), Hes1 +/− :p107 −/− ( n = 3), and p107 −/− ( n = 3) brains. (c) A 2-h BrdU pulse labeled proliferating progenitors in E13.5 brains. BrdU-positive cells were counted in four representative regions of the brain in wild-type ( n = 4), Hes1 +/− :p107 −/− ( n = 4), Hes1 +/− ( n = 4), and p107 −/− ( n = 3) embryos. Note that loss of a single Hes1 allele restored the numbers of progenitor cells to wild-type levels in p107 −/− mice at embryonic and adult ages. Means were statistically analyzed by one-way analysis of variance followed by Tukey's individual comparison of the means. Error bars represent SEM. *, P

    Article Snippet: Immunohistochemistry and Western blotting Immunohistochemistry was performed on coronal cryostat sections from embryonic and adult brains with primary antibodies to mouse anti-NeuN (1:100; Chemicon), rabbit antiactive caspase-3 (1:500; BD Biosciences), rabbit anti-PH3 (1:400; Upstate Biotechnology), rat anti-BrdU (1:100; Accurate Chemicals), mouse anti-BrdU (1:100; Becton Dickinson), mouse anti-PCNA (1:300; Vector Laboratories), goat antidoublecortin (1:100; Santa Cruz Biotechnology, Inc.), mouse anti–βIII-tubulin (mouse monoclonal hybridoma supernatant; 1:100; ), and mouse anti-Nestin (1:400; Research Diagnostics).

    Techniques: Mouse Assay, Immunohistochemistry, Labeling

    Increased numbers of progenitor cells in the embryonic and adult p107 −/− brains in vivo. (a–c) To assess the size of the neural progenitor population in the adult brain, mice received intraperitoneal injections of BrdU to label proliferating cells over a 10-h period. BrdU-positive cell counts through the lateral ventricles showed a 50% increase in the number of progenitors in p107 −/− brains ( n = 4) in comparison with wild type ( n = 3). (d) The adult progenitor population was also assessed by immunohistochemistry for PCNA. Similar to our BrdU incorporation study, p107-deficient brains had a comparable increase in PCNA + cells along the ventricles. (e) To quantify the number of proliferating progenitors in E10.5 brains, we used an antibody to phosphohistone H3 (PH3), a marker of cells in M phase of the cell cycle. PH3-positive cells were counted along the lateral ventricles in three representative sections of wild-type ( n = 3) and p107 −/− ( n = 3) brains. (f) A 2-h BrdU pulse was used to label progenitors in S phase in E13.5 brains. BrdU-positive cells were counted in four representative regions of the forebrain in wild-type ( n = 4) and p107 −/− ( n = 3) embryos. Note that at adult and embryonic ages, p107 −/− mice had significantly more proliferating cells along the lateral ventricle than wild-type mice. (g and h) To assess whether in vivo cell cycle kinetics were different in p107 −/− neural progenitors, we compared the size of the S-phase fraction by immunolabeling for BrdU and PCNA in adult (g) and E13.5 wild-type and p107 −/− cortices (h). Quantification of BrdU+ cells (S-phase fraction) was compared with the total proliferating population (PCNA + ). After 10.5 h of cumulative BrdU labeling, 78.2% and 78% of the total proliferating populations in wild-type and p107 −/− cortices were BrdU + , respectively, indicating similar cell cycle kinetics. At E13.5 after a 2-h BrdU pulse, 53.6% and 51.2% of the total proliferating populations in wild-type and p107 −/− were BrdU + , respectively. Means were statistically analyzed using a t test, and differences were assessed at *, P

    Journal: The Journal of Cell Biology

    Article Title: The Retinoblastoma family member p107 regulates the rate of progenitor commitment to a neuronal fate

    doi: 10.1083/jcb.200703176

    Figure Lengend Snippet: Increased numbers of progenitor cells in the embryonic and adult p107 −/− brains in vivo. (a–c) To assess the size of the neural progenitor population in the adult brain, mice received intraperitoneal injections of BrdU to label proliferating cells over a 10-h period. BrdU-positive cell counts through the lateral ventricles showed a 50% increase in the number of progenitors in p107 −/− brains ( n = 4) in comparison with wild type ( n = 3). (d) The adult progenitor population was also assessed by immunohistochemistry for PCNA. Similar to our BrdU incorporation study, p107-deficient brains had a comparable increase in PCNA + cells along the ventricles. (e) To quantify the number of proliferating progenitors in E10.5 brains, we used an antibody to phosphohistone H3 (PH3), a marker of cells in M phase of the cell cycle. PH3-positive cells were counted along the lateral ventricles in three representative sections of wild-type ( n = 3) and p107 −/− ( n = 3) brains. (f) A 2-h BrdU pulse was used to label progenitors in S phase in E13.5 brains. BrdU-positive cells were counted in four representative regions of the forebrain in wild-type ( n = 4) and p107 −/− ( n = 3) embryos. Note that at adult and embryonic ages, p107 −/− mice had significantly more proliferating cells along the lateral ventricle than wild-type mice. (g and h) To assess whether in vivo cell cycle kinetics were different in p107 −/− neural progenitors, we compared the size of the S-phase fraction by immunolabeling for BrdU and PCNA in adult (g) and E13.5 wild-type and p107 −/− cortices (h). Quantification of BrdU+ cells (S-phase fraction) was compared with the total proliferating population (PCNA + ). After 10.5 h of cumulative BrdU labeling, 78.2% and 78% of the total proliferating populations in wild-type and p107 −/− cortices were BrdU + , respectively, indicating similar cell cycle kinetics. At E13.5 after a 2-h BrdU pulse, 53.6% and 51.2% of the total proliferating populations in wild-type and p107 −/− were BrdU + , respectively. Means were statistically analyzed using a t test, and differences were assessed at *, P

    Article Snippet: Immunohistochemistry and Western blotting Immunohistochemistry was performed on coronal cryostat sections from embryonic and adult brains with primary antibodies to mouse anti-NeuN (1:100; Chemicon), rabbit antiactive caspase-3 (1:500; BD Biosciences), rabbit anti-PH3 (1:400; Upstate Biotechnology), rat anti-BrdU (1:100; Accurate Chemicals), mouse anti-BrdU (1:100; Becton Dickinson), mouse anti-PCNA (1:300; Vector Laboratories), goat antidoublecortin (1:100; Santa Cruz Biotechnology, Inc.), mouse anti–βIII-tubulin (mouse monoclonal hybridoma supernatant; 1:100; ), and mouse anti-Nestin (1:400; Research Diagnostics).

    Techniques: In Vivo, Mouse Assay, Immunohistochemistry, BrdU Incorporation Assay, Marker, Immunolabeling, Labeling