anti-phospho-histone h3 Search Results


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  • 94
    Thermo Fisher anti phospho histone h3
    VEGF signaling mediated by VEGFR2 is not required for cell proliferation, cell survival, expression of FoxP1, or nuclear translocation of NFATc1 in endocardial cells of elongating mitral valves (MVs). (A, B) Immunofluorescent staining for <t>phospho-histone</t>
    Anti Phospho Histone H3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti phospho histone h 3
    A: An experimental strategy for transcriptomic analysis of dissected midguts from flies with modified IMD activity. B: Principle component analysis of gene expression data for control flies ( Myo1A ts / + ), and flies with IMD activity blocked in enterocytes ( Myo1A ts / D30A ). C: Volcano plot of genes that are differentially expressed in  Myo1A ts / D30A  midguts relative to  Myo1A ts / +  midguts. Each point represents a single gene. Orange indicates genes with a greater than 2 fold-change in gene expression. Green indicates genes with a greater than 2 fold-change in gene expression, and an FDR below 0.01. D: Gene Ontology term analysis of pathways modified by inhibition of IMD in enterocytes. Column size indicates fold-enrichment, and circles show the significance of that enrichment on a negative log scale. E-J: Quantification of glucose (E-F), trehalose (G), triglyceride (H, I), and weight (J) of whole flies, or dissected midguts from flies of the indicated genotypes. P values are from significance tests performed with Student’s t-tests for each measurement. K: Quantification of phospho-histone H3-positive mitotic cells in the posterior midguts of flies of the indicated genotypes raised at 29°C for 28 days. L: Survival curves of control flies (-), or flies with IMD activity blocked in enterocytes (+). N=number of flies for each genotype. Chi-squared and P values are from Log-rank tests.
    Anti Phospho Histone H 3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho histone h3
    Translational blockade of zebrafish  shox  reduced cell proliferation and specific gene expression of skeletal progenitors in developing embryos.  (A)  Analyses of cell proliferation and cell death of  shox  morpholino oligo (MO) or control MO ( ctr  MO)-injected 22 hpf embryos. Bars, 100 µm. Representative results of phospho-Histone H3-staining and active-caspase-3-staining. Quantification data of phospho-Histone H3-staining of ctr MO-injected embryos ( n  = 5) and shox MO-injected embryos ( n  = 9). * P
    Anti Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 510 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho histone h3
    Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated <t>histone</t> H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.
    Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1255 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho histone h3 s10
    Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated <t>histone</t> H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.
    Anti Phospho Histone H3 S10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho histone h3 thr11
    Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated <t>histone</t> H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.
    Anti Phospho Histone H3 Thr11, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho histone h3 ser28
    Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated <t>histone</t> H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.
    Anti Phospho Histone H3 Ser28, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti phospho histone h3 antibody
    Generation and characterization of cardiac myocyte-specific Hdac3-Tg mice. A , schematic of transgenic construct used to generate Hdac3 -Tg mice. α -MHC , α-myosin heavy chain. B , Hdac3 mRNA expression analysis in two different Hdac3 -Tg mice lines. Transcripts for Hdac3 were detected by qRT-PCR in P1 hearts from wild-type ( WT ) and Hdac3 -Tg mice. Three mice in each group were tested, and values are expressed as the -fold change in transcript abundance (± S.D.) when compared with wild-type mice. C , endogenous and transgenic Hdac3 protein were detected with an Hdac3 antibody. All Western blots were performed at least three times with similar results. D , Hdac3 expression in myocardium of P1 wild-type and Hdac3 -Tg mice was quantified by using ImageJ software. E , increased HDAC activity in Hdac3 -Tg mice. Lysates from P1 hearts were assayed for HDAC activity. Data are the average results (± S.D.) from three separate experiments. F , decreased acetylation of <t>histone</t> H4 in Hdac3 -Tg mice. Western blot analysis of acetylated-histone H4 in myocardium from P1 wild-type and Hdac3 -Tg mice using anti-acetyl-histone H4 antibody was performed. ImageJ software was used to quantify -fold change in acetylation. G , Hdac1 and Hdac2 levels are not changed in Hdac3 -Tg mice. Western blot analysis of myocardium lysates from three P1 wild-type and three Hdac3 -Tg mice using anti-Hdac1 and anti-Hdac2 antibody was performed. α-Tubulin is used as a loading control.
    Anti Phospho Histone H3 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Upstate Biotechnology Inc anti phospho histone h3
    Proliferation and initial differentiation of interneuron precursors in the PWM. a–c , Distribution of dividing interneuron precursors in Pax-2-GFP transgenic mice at three different postnatal ages (P2, P6, P9). a , b , Graphs illustrate the frequency of GFP/BrdU double-labeled cells in PWM and cortical layers, 2 h ( a ) or 24 h ( b ) after BrdU administration ( n = 2–4 cases/time point). c–e , The majority of dividing cells are located in the PWM (arrows in c ). This distribution is confirmed by immunostaining for <t>phospho-histone</t> Ph3 ( d , e ). f , Laminar position of GFP/BrdU-double-labeled cells at increasing times after BrdU injection: 4 d after nucleotide analog administration, many cells are still in the PWM. g–n shows Pax-2 immunolabeling ( g–j ) or Pax-2/GFP expression ( k–n ) in interneuron precursors labeled by BrdU 2 h ( g , h , k , l ) or 24 h ( i , j , m , n ) before animals were killed. o , p , Double-labeled cells (arrows) consistently show a weaker Pax-2 expression at 2 h than at 24 h, as confirmed by the quantitative estimation of Pax-2 immunofluorescence intensity ( o , Student's t test, p
    Anti Phospho Histone H3, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 241 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck KGaA anti phospho histone h3
    Overexpression of cyclin B1 reduces the survival of BRCA1-deficient cells. a  Unsynchronized MCF7 cells in which BRCA1 was knocked down and/or cyclin B1 was overexpressed were examined according to the cell cycle phase, determined by flow cytometry. Representative histograms showing the DNA content in MCF7 cells with BRCA1 depletion and/or cyclin B1 overexpression.  b  Percentages of cells in each cell cycle phase are shown.  c  Protein expression patterns in  BRCA1 -knockdown and/or cyclin B1-overexpressing MCF7 cells were analyzed via Western blot analysis.  d  To count the cells in G2/M transition, unsynchronized MCF7 cells transfected with BRCA1 siRNA and/or transduced with Ad-cyclin B1 were stained with phospho (Ser10)-Histone H3 and analyzed via flow cytometry.  e  Survival of cyclin B1-overexpressing MCF7 cells was measured in cells transfected with scrambled siRNA or  BRCA1 -targeted siRNA. The results are expressed as survival relative to that at infection or transfection.  f  A breast cancer cell line derived from a  Brca1 Δ11/Δ11 p53 −/−  mouse was infected with increasing amounts of Ad-cyclin B1, after which survival was analyzed with an MTT assay.  g  A breast cancer cell line derived from a  Brca1 Δ11/Δ11 p53 −/−  mouse was infected with Ad-cyclin B1 or Ad-GFP (control), after which the protein pattern was analyzed via Western blotting.  h  Estimated survival of  Brca1 Δ11/Δ11 ;p53 −/−  mammary tumor cell lines at the indicated concentration of vinblastine.  i  The survival of  Brca1 Δ11/Δ11 p53 −/−  mammary tumor cell lines was assessed after transfection with scramble or  Ccnb1  siRNA and treatment with vinblastine at the indicated concentration. ** P
    Anti Phospho Histone H3, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti phospho histone h3
    Lentiviral knockdown of FoxF1, FoxF2 or both in mouse and human RMS cells decreased cellular growth  in vitro (A)  Efficiency of FoxF1 and FoxF2 lentiviral knockdown in mouse 76-9 rhabdomyosarcoma cells was shown by qRT-PCR.  β-actin  mRNA was used for normalization. ( B)  Depletion of FoxF1, FoxF2 or both decreased proliferation of 76-9 cells  in vitro . Control, FoxF1-KD, FoxF2-KD and F1+F2-KD 76-9 cells were seeded in triplicates and counted at different time points using WST1 Cell Proliferation Assay. ( C)  Depletion of FoxF1, FoxF2 or both decreased colony formation in soft agar compared to control RMS cells. Cells were seeded in triplicates. Values are the means ± SD of three independent experiments. A  p value  lt;0.05 is shown with (*). ( D)  Depletion of FoxF1, FoxF2 or both decreased the number of cells in mitosis shown by flow cytometery with phospho-Histone H3 antibodies.  (E)  Efficiency of FoxF1 and FoxF2 lentiviral knockdown in human Rh30 rhabdomyosarcoma cells was shown by qRT-PCR.  β-actin  mRNA was used for normalization. ( F)  Depletion of FoxF1, FoxF2 or both decreased proliferation of Rh30 cells  in vitro . Control, FoxF1-, FoxF2- or F1+F2-deficient Rh30 cells were seeded in triplicates and counted at different time points using hemocytometer. Data represent mean±s.d. of three independent experiments. A  p  value   lt; 0.05 is shown with (*);  p  value   lt; 0.01 is shown with (**).
    Anti Phospho Histone H3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti phospho histone h3
    SHP-2 N308D leads to a delay of the cardiac cell cycle. ( A-D ) Transverse heart sections through stage 33 (A,B) and stage 37 (C,D) Xenopus embryos stained with Tmy (green) and anti-phospho <t>histone</t> H3 (red), from uninjected (A,C) and SHP-2 N308D -derived (B,D)
    Rabbit Anti Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    VEGF signaling mediated by VEGFR2 is not required for cell proliferation, cell survival, expression of FoxP1, or nuclear translocation of NFATc1 in endocardial cells of elongating mitral valves (MVs). (A, B) Immunofluorescent staining for phospho-histone

    Journal: Developmental biology

    Article Title: VEGF Signaling has Distinct Spatiotemporal Roles During Heart Valve Development

    doi: 10.1016/j.ydbio.2010.08.030

    Figure Lengend Snippet: VEGF signaling mediated by VEGFR2 is not required for cell proliferation, cell survival, expression of FoxP1, or nuclear translocation of NFATc1 in endocardial cells of elongating mitral valves (MVs). (A, B) Immunofluorescent staining for phospho-histone

    Article Snippet: For anti-PECAM (BD Biosciences, San Jose, CA), anti-smooth muscle actin (Sigma, St. Louis, MO), and anti-phospho-histone H3 (Upstate, Billerica, MA), sections were exposed to 0.05% trypsin (Invitrogen, Carlsbad, CA) for 45 minutes at 37°C.

    Techniques: Expressing, Translocation Assay, Staining

    A: An experimental strategy for transcriptomic analysis of dissected midguts from flies with modified IMD activity. B: Principle component analysis of gene expression data for control flies ( Myo1A ts / + ), and flies with IMD activity blocked in enterocytes ( Myo1A ts / D30A ). C: Volcano plot of genes that are differentially expressed in  Myo1A ts / D30A  midguts relative to  Myo1A ts / +  midguts. Each point represents a single gene. Orange indicates genes with a greater than 2 fold-change in gene expression. Green indicates genes with a greater than 2 fold-change in gene expression, and an FDR below 0.01. D: Gene Ontology term analysis of pathways modified by inhibition of IMD in enterocytes. Column size indicates fold-enrichment, and circles show the significance of that enrichment on a negative log scale. E-J: Quantification of glucose (E-F), trehalose (G), triglyceride (H, I), and weight (J) of whole flies, or dissected midguts from flies of the indicated genotypes. P values are from significance tests performed with Student’s t-tests for each measurement. K: Quantification of phospho-histone H3-positive mitotic cells in the posterior midguts of flies of the indicated genotypes raised at 29°C for 28 days. L: Survival curves of control flies (-), or flies with IMD activity blocked in enterocytes (+). N=number of flies for each genotype. Chi-squared and P values are from Log-rank tests.

    Journal: bioRxiv

    Article Title: Cell-Specific Regulation of Intestinal Immunity

    doi: 10.1101/721662

    Figure Lengend Snippet: A: An experimental strategy for transcriptomic analysis of dissected midguts from flies with modified IMD activity. B: Principle component analysis of gene expression data for control flies ( Myo1A ts / + ), and flies with IMD activity blocked in enterocytes ( Myo1A ts / D30A ). C: Volcano plot of genes that are differentially expressed in Myo1A ts / D30A midguts relative to Myo1A ts / + midguts. Each point represents a single gene. Orange indicates genes with a greater than 2 fold-change in gene expression. Green indicates genes with a greater than 2 fold-change in gene expression, and an FDR below 0.01. D: Gene Ontology term analysis of pathways modified by inhibition of IMD in enterocytes. Column size indicates fold-enrichment, and circles show the significance of that enrichment on a negative log scale. E-J: Quantification of glucose (E-F), trehalose (G), triglyceride (H, I), and weight (J) of whole flies, or dissected midguts from flies of the indicated genotypes. P values are from significance tests performed with Student’s t-tests for each measurement. K: Quantification of phospho-histone H3-positive mitotic cells in the posterior midguts of flies of the indicated genotypes raised at 29°C for 28 days. L: Survival curves of control flies (-), or flies with IMD activity blocked in enterocytes (+). N=number of flies for each genotype. Chi-squared and P values are from Log-rank tests.

    Article Snippet: In brief, we used anti-phospho-histone H3 (PH3, 1:1000, Millipore (Upstate), 06-570) immunofluorescence to quantify mitoses in the midguts, and anti-Delta (1:100; Developmental Studies Hybridoma Bank C594.9B) immunofluorescence to quantify stem cells in the R4/R5 region of the posterior midguts of virgin female flies that we raised at 29°C for 5d or 30d.

    Techniques: Modification, Radial Immuno Diffusion, Activity Assay, Expressing, Inhibition

    A: Percentage of cells positive for the stem cell marker, Delta in the posterior midgut of flies of the indicated genotypes and ages. P value are from a significance test performed with a Student’s t-test. B: Quantification of phospho-histone H3-positive mitotic cells in the posterior midguts of 28d old flies of the indicated genotypes. P value are from a significance test performed with a Student’s t-test. C-D: Survival curves of female control flies (-), or flies with IMD activity blocked in the progenitors (+). N=number of flies tested for each genotype. Chi-squared and P values are the results of Log-rank tests. Survival studies were performed with two distinct  imdD30A -expressing flies (labeled 1 and 2, respectively).

    Journal: bioRxiv

    Article Title: Cell-Specific Regulation of Intestinal Immunity

    doi: 10.1101/721662

    Figure Lengend Snippet: A: Percentage of cells positive for the stem cell marker, Delta in the posterior midgut of flies of the indicated genotypes and ages. P value are from a significance test performed with a Student’s t-test. B: Quantification of phospho-histone H3-positive mitotic cells in the posterior midguts of 28d old flies of the indicated genotypes. P value are from a significance test performed with a Student’s t-test. C-D: Survival curves of female control flies (-), or flies with IMD activity blocked in the progenitors (+). N=number of flies tested for each genotype. Chi-squared and P values are the results of Log-rank tests. Survival studies were performed with two distinct imdD30A -expressing flies (labeled 1 and 2, respectively).

    Article Snippet: In brief, we used anti-phospho-histone H3 (PH3, 1:1000, Millipore (Upstate), 06-570) immunofluorescence to quantify mitoses in the midguts, and anti-Delta (1:100; Developmental Studies Hybridoma Bank C594.9B) immunofluorescence to quantify stem cells in the R4/R5 region of the posterior midguts of virgin female flies that we raised at 29°C for 5d or 30d.

    Techniques: Marker, Radial Immuno Diffusion, Activity Assay, Expressing, Labeling

    Cilia effects on proliferation and ECM production ( A ) IHC of postnatal day 0 (P0) aortic valves, for proliferation markers Ki67 and phospho-histone H3, show no difference in proliferating or total cell number when conditional knockout aortic valves were compared to littermate controls, quantified in ( B ). ( C ) H and E staining’s show increased matrix in the fused right-non-coronary leaflet of the conditional knockout. C′= wild-type right coronary, C″= wild-type right coronary tip, C*= conditional knockout right non-coronary base, C**= conditional knockout right coronary tip. RC= right coronary, LC= left coronary, RNC= right non-coronary. ( D ) Quantification of cell density shows a significant decrease in cell density in cilia deficient valves at both the base and tip of the right coronary leaflet, with p

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    Article Title: A Role for Primary Cilia in Aortic Valve Development and Disease

    doi: 10.1002/dvdy.24524

    Figure Lengend Snippet: Cilia effects on proliferation and ECM production ( A ) IHC of postnatal day 0 (P0) aortic valves, for proliferation markers Ki67 and phospho-histone H3, show no difference in proliferating or total cell number when conditional knockout aortic valves were compared to littermate controls, quantified in ( B ). ( C ) H and E staining’s show increased matrix in the fused right-non-coronary leaflet of the conditional knockout. C′= wild-type right coronary, C″= wild-type right coronary tip, C*= conditional knockout right non-coronary base, C**= conditional knockout right coronary tip. RC= right coronary, LC= left coronary, RNC= right non-coronary. ( D ) Quantification of cell density shows a significant decrease in cell density in cilia deficient valves at both the base and tip of the right coronary leaflet, with p

    Article Snippet: The following are the antibodies and their dilutions; Acetylated Tubulin (Sigma, Cat#T6793, 1:500), Gamma Tubulin (Abcam, Cambridge, MA, USA, Cat#ab11317, 1:1000), Versican (gift from Stan Hoffman, Medical University of South Carolina, 1: 250), Collagen (gift from Stan Hoffman, Medical University of South Carolina, 1: 250), MF20 (DSHB, Iowa City, IA, USA, Concentrate, I:50), Ki67 (Abcam, Cat#ab16667, 1:250), Phospho-histone H3 (EMD Millipore, Darmstadt Germany, Cat#06-570, 1:250), Smoothened (LSBio, Seattle WA, Cat#LS-A2666, 1:250), Gli3 (Origene, Rockville MD, Cat#TA337186, 1:250).

    Techniques: Immunohistochemistry, Knock-Out, Staining

    Pdlim7 knock-down pectoral fins have decreased cell proliferation . A-F: Dorsal view, anterior end of embryo is out of view to the bottom of image, of whole-mount anti-phospho-histone H3 (p-H3) antibody (red) staining on wild-type (A, C, E) and MO2 injected (B, D, F) embryos. Pectoral fins develop lateral to the 3 rd  somite, thus embryos were counterstained with MF20 (blue) to visualize somites, which are indicated by numbers (somite 1 refers to the most anterior somite). G: Quantification of p-H3 positive cells in pectoral fins of wild-type and MO2 injected embryos. 28 hpf wild-type n = 5, MO2 n = 5; p-value = 0.077. 36 hpf wild-type n = 4, MO2 n = 5; p-value = 0.036. 48 hpf wild-type n = 5, MO2 n = 5; p-value = 0.001. Experiment performed in triplicate, representative data from single replicate shown in G. Statistically significant p-values (

    Journal: BMC Developmental Biology

    Article Title: Pdlim7 is required for maintenance of the mesenchymal/epidermal Fgf signaling feedback loop during zebrafish pectoral fin development

    doi: 10.1186/1471-213X-10-104

    Figure Lengend Snippet: Pdlim7 knock-down pectoral fins have decreased cell proliferation . A-F: Dorsal view, anterior end of embryo is out of view to the bottom of image, of whole-mount anti-phospho-histone H3 (p-H3) antibody (red) staining on wild-type (A, C, E) and MO2 injected (B, D, F) embryos. Pectoral fins develop lateral to the 3 rd somite, thus embryos were counterstained with MF20 (blue) to visualize somites, which are indicated by numbers (somite 1 refers to the most anterior somite). G: Quantification of p-H3 positive cells in pectoral fins of wild-type and MO2 injected embryos. 28 hpf wild-type n = 5, MO2 n = 5; p-value = 0.077. 36 hpf wild-type n = 4, MO2 n = 5; p-value = 0.036. 48 hpf wild-type n = 5, MO2 n = 5; p-value = 0.001. Experiment performed in triplicate, representative data from single replicate shown in G. Statistically significant p-values (

    Article Snippet: Next, the Anti-phospho-histone-h3 antibody (Anti-p-H3) (Millipore) was used at a 1:20 dilution, while the MF20 antibody was used at a 1:40 dilution in PBT with 0.2% saponin.

    Techniques: Staining, Injection

    Loss of  ftr82  impairs the growth of ISV cells. ( A , A’ , B , B’ ) TdT-mediated dUTP-X nick end labeling (TUNEL) assay was used to detect apoptotic cells in uninjected control and  ftr82 e1i1  morphants. Increased apoptotic cells were observed on the skin and in the epidermis of the dorsal tail region (arrows), but not in vascular regions in  ftr82  MO compared to controls at 28 hpf; ( A’ , B’ ) are expanded images of ( A ) and ( B ), respectively; ( C , D ) AO staining (green dots) in  Tg  ( kdrl:mCherry ) fish exhibited more apoptotic cells with the knockdown of  ftr82 ; ( E , F ) The number of cells forming each ISV counted in control  Tg  ( kdrl:mCherry ci5 ;  fli1a:negfp  y7 ) ( E ) and  ftr82  morphant embryos ( F ) at 32 hpf; ( G ) Quantification of average ISV cells per ISV counted in both control ( n  = 16) and  ftr82  morphant ( n  = 18); ( H – L ) migration assay measured the difference of ISV length from 24 to 28 hpf in control and  ftr82  MO ( n  = 30 ISVs from three control embryos or morphants); ( M , N ) Proliferation marker pHH3 was counted in the trunk region beneath the neural tube and above the yolk extension area, where it is more related to main vessels and ISVs; ( O ) The mean number of mitotic cells (phosphohistone H3, pHH3 cells) in control was 16.3 ± 2.4 ( n  = 8) and in  ftr82  MO was 7.5 ± 2.1 ( n  = 8); ( P ) Western blot analysis showed the reduced expression level of pHH3; the increase of p21 and p27 protein levels. β-actin serves as a loading control. Scale bars are 200 μm for ( A – D , A’ , B’ , M , N ) and 50 μm for ( E , F , H – K ). *** refers to  p

    Journal: International Journal of Molecular Sciences

    Article Title: Ftr82 Is Critical for Vascular Patterning during Zebrafish Development

    doi: 10.3390/ijms18010156

    Figure Lengend Snippet: Loss of ftr82 impairs the growth of ISV cells. ( A , A’ , B , B’ ) TdT-mediated dUTP-X nick end labeling (TUNEL) assay was used to detect apoptotic cells in uninjected control and ftr82 e1i1 morphants. Increased apoptotic cells were observed on the skin and in the epidermis of the dorsal tail region (arrows), but not in vascular regions in ftr82 MO compared to controls at 28 hpf; ( A’ , B’ ) are expanded images of ( A ) and ( B ), respectively; ( C , D ) AO staining (green dots) in Tg ( kdrl:mCherry ) fish exhibited more apoptotic cells with the knockdown of ftr82 ; ( E , F ) The number of cells forming each ISV counted in control Tg ( kdrl:mCherry ci5 ; fli1a:negfp y7 ) ( E ) and ftr82 morphant embryos ( F ) at 32 hpf; ( G ) Quantification of average ISV cells per ISV counted in both control ( n = 16) and ftr82 morphant ( n = 18); ( H – L ) migration assay measured the difference of ISV length from 24 to 28 hpf in control and ftr82 MO ( n = 30 ISVs from three control embryos or morphants); ( M , N ) Proliferation marker pHH3 was counted in the trunk region beneath the neural tube and above the yolk extension area, where it is more related to main vessels and ISVs; ( O ) The mean number of mitotic cells (phosphohistone H3, pHH3 cells) in control was 16.3 ± 2.4 ( n = 8) and in ftr82 MO was 7.5 ± 2.1 ( n = 8); ( P ) Western blot analysis showed the reduced expression level of pHH3; the increase of p21 and p27 protein levels. β-actin serves as a loading control. Scale bars are 200 μm for ( A – D , A’ , B’ , M , N ) and 50 μm for ( E , F , H – K ). *** refers to p

    Article Snippet: The membranes were washed in TBS (Tris-buffered saline) with Tween-20 (TBST) and blocked with nonfat milk in TBST for 1 h. The membranes were then incubated with anti-phospho histone H3 (pHH3) (Millipore, Billerica, MA, USA), anti-p21, anti-p27 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) polyclonal antibodies, and anti-β-actin monoclonal antibody (Sigma) overnight at 4 °C.

    Techniques: End Labeling, TUNEL Assay, Staining, Fluorescence In Situ Hybridization, Migration, Marker, Western Blot, Expressing

    Western blots, showing GFP-aurC and GFP-alone proteins after 24 hours of transient transfection with GFP-alone, GFP-aurC-KD GFP-aurC-CA and GFP-aurC-WT plasmid DNA with mouse Anti-GFP antibody (A) and with rabbit Anti-aurC antibody (B) . Western blots showing the level of expression of GFP-aurC protein in three stable clones of GFP-aurC-KD (KD1 to KD3), four stable clones of GFP-aurC-CA (CA1 to CA4), three stable clones each of GFP-aurC-WT (WT1 to WT3) and GFP-alone (GFP1 to GFP3) illustrating the different level of expression of GFP-aurC and GFP proteins by different clones. The antibody used was mouse anti-GFP ( C   D ) and anti-β tubulin antibody as a loading control ( E    F ); ( G ) Kinase assay GFP-aurC-WT, GFP-aurC-CA and GFP-alone clones, using histone-H3 as a substrate. ( H 1,2,3,4 ) The left column shows DAPI stained cells and the right column shows phosphorylated cells with Histone-H3 ser-10. ( H-1 ) GFP-aurC-WT and ( H-2 ) GFP-aurC-CA ( H-3 ) GFP-aurC-KD.  (H-4)  GFP-alone ( I ) Histogram shows the percentage of cells with phosphorylation on histone H3 of GFP-aurC-WT, GFP-aurC-CA, GFP-aurC-KD and GFP-alone.

    Journal: BMC Cell Biology

    Article Title: Aurora kinase-C-T191D is constitutively active mutant

    doi: 10.1186/1471-2121-13-8

    Figure Lengend Snippet: Western blots, showing GFP-aurC and GFP-alone proteins after 24 hours of transient transfection with GFP-alone, GFP-aurC-KD GFP-aurC-CA and GFP-aurC-WT plasmid DNA with mouse Anti-GFP antibody (A) and with rabbit Anti-aurC antibody (B) . Western blots showing the level of expression of GFP-aurC protein in three stable clones of GFP-aurC-KD (KD1 to KD3), four stable clones of GFP-aurC-CA (CA1 to CA4), three stable clones each of GFP-aurC-WT (WT1 to WT3) and GFP-alone (GFP1 to GFP3) illustrating the different level of expression of GFP-aurC and GFP proteins by different clones. The antibody used was mouse anti-GFP ( C D ) and anti-β tubulin antibody as a loading control ( E F ); ( G ) Kinase assay GFP-aurC-WT, GFP-aurC-CA and GFP-alone clones, using histone-H3 as a substrate. ( H 1,2,3,4 ) The left column shows DAPI stained cells and the right column shows phosphorylated cells with Histone-H3 ser-10. ( H-1 ) GFP-aurC-WT and ( H-2 ) GFP-aurC-CA ( H-3 ) GFP-aurC-KD. (H-4) GFP-alone ( I ) Histogram shows the percentage of cells with phosphorylation on histone H3 of GFP-aurC-WT, GFP-aurC-CA, GFP-aurC-KD and GFP-alone.

    Article Snippet: Immunohistochemistry was performed with rabbit monoclonal KI-67 (1.200, Epitomics, clone SP6) and anti-phospho histone- H3 ser-10 (Millipore USA) and anti-HRP (Jackson USA) secondary antibodies.

    Techniques: Western Blot, Transfection, Plasmid Preparation, Expressing, Clone Assay, Kinase Assay, Staining

    Soft agar assay, tumour formation and immunohistochemistry .  (A)  Foci of colonies of GFP-aurA, GFP-aurC-CA, GFP-aurC-WT stable cell lines and very negligible number of very small colonies of GFP-alone in soft agar assay.  (B)  Histogram of the average number of colonies.  (C)  Visualization of the tumours formed by injecting GFP-aurC-CA and GFP-aurC-WT stable cell lines, and the remaining injected cells of GFP-alone on the day of sacrifice using Kodak image station 2000.  (D)  Rabbit monoclonal KI-67, a proliferation marker from late G 1  to M-phase staining  (E)  anti-phospho histone-H3 ser-10 (Millipore) and anti-HRP (Jackson) secondary antibodies and  (F    G)  Feulgen staining showing prometaphase defects, metaphase defects, lagging chromosomes at anaphase, and cytokinesis defect.

    Journal: BMC Cell Biology

    Article Title: Aurora kinase-C-T191D is constitutively active mutant

    doi: 10.1186/1471-2121-13-8

    Figure Lengend Snippet: Soft agar assay, tumour formation and immunohistochemistry . (A) Foci of colonies of GFP-aurA, GFP-aurC-CA, GFP-aurC-WT stable cell lines and very negligible number of very small colonies of GFP-alone in soft agar assay. (B) Histogram of the average number of colonies. (C) Visualization of the tumours formed by injecting GFP-aurC-CA and GFP-aurC-WT stable cell lines, and the remaining injected cells of GFP-alone on the day of sacrifice using Kodak image station 2000. (D) Rabbit monoclonal KI-67, a proliferation marker from late G 1 to M-phase staining (E) anti-phospho histone-H3 ser-10 (Millipore) and anti-HRP (Jackson) secondary antibodies and (F G) Feulgen staining showing prometaphase defects, metaphase defects, lagging chromosomes at anaphase, and cytokinesis defect.

    Article Snippet: Immunohistochemistry was performed with rabbit monoclonal KI-67 (1.200, Epitomics, clone SP6) and anti-phospho histone- H3 ser-10 (Millipore USA) and anti-HRP (Jackson USA) secondary antibodies.

    Techniques: Soft Agar Assay, Immunohistochemistry, Stable Transfection, Injection, Marker, Staining

    Translational blockade of zebrafish  shox  reduced cell proliferation and specific gene expression of skeletal progenitors in developing embryos.  (A)  Analyses of cell proliferation and cell death of  shox  morpholino oligo (MO) or control MO ( ctr  MO)-injected 22 hpf embryos. Bars, 100 µm. Representative results of phospho-Histone H3-staining and active-caspase-3-staining. Quantification data of phospho-Histone H3-staining of ctr MO-injected embryos ( n  = 5) and shox MO-injected embryos ( n  = 9). * P

    Journal: Frontiers in Endocrinology

    Article Title: The Short-Stature Homeobox-Containing Gene (shox/SHOX) Is Required for the Regulation of Cell Proliferation and Bone Differentiation in Zebrafish Embryo and Human Mesenchymal Stem Cells

    doi: 10.3389/fendo.2017.00125

    Figure Lengend Snippet: Translational blockade of zebrafish shox reduced cell proliferation and specific gene expression of skeletal progenitors in developing embryos. (A) Analyses of cell proliferation and cell death of shox morpholino oligo (MO) or control MO ( ctr MO)-injected 22 hpf embryos. Bars, 100 µm. Representative results of phospho-Histone H3-staining and active-caspase-3-staining. Quantification data of phospho-Histone H3-staining of ctr MO-injected embryos ( n  = 5) and shox MO-injected embryos ( n  = 9). * P

    Article Snippet: The whole-mount immunostaining was conducted using anti-phospho-Histone H3 (3377, Cell Signaling Technology Japan, Tokyo, Japan), anti-active-caspase-3 (559565, BD Pharmingen™, BD Biosciences Japan, Tokyo, Japan) and anti-myosin heavy chain (05-716, Millipore, Darmstadt, Germany) according to the instruction manuals.

    Techniques: Expressing, Injection, Staining

    Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated histone H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Fatty Acid Oxidation Promotes Cardiomyocyte Proliferation Rate but Does Not Change Cardiomyocyte Number in Infant Mice

    doi: 10.3389/fcell.2019.00042

    Figure Lengend Snippet: Cardiomyocyte cycling and metabolic profiling in infant mouse cardiomyocytes. (A) Isolated cardiomyocytes in DNA synthesis-phase were visualized by immunofluorescent microscopy using Click-iT EdU Alexa Fluor (red) and co-immunostaining with antibody against cardiac troponin T (cTnT, green). Arrows point to EdU+cTnT+ cells. Arrowheads point to binuclear cTnT+ cells. (B) Quantification of EdU+cTnT+ cells as percentage of total cTnT+ cells (∼1200 cTnT+ cells per sample). (C) Confocal images cardiomyocytes in mitotic phase as detected by co-immunostainings for phosphorylated histone H3 (PH3, red) and cTnT (green) on tissue sections, and quantification of PH3+cTnT+ cells as percentage of total cTnT+ cells analyzed per field. Arrows point to PH3+cTnT+ cells. (D–G) Isolated mouse cardiomyocytes from postnatal day 2 (P2), P5 and P7 heart ventricles were assessed with the Seahorse XF Analyzer. (D) Measurement and (E) quantification of mitochondrial oxygen consumption rate (OCR) with fatty acid stress test using palmitate versus BSA control. (F) Measurement and (G) quantification of extracellular acidification rate (ECAR) in the glycolysis stress assay. 2-DG (2-deoxyglucose) is a hexokinase inhibitor, which inhibits glycolytic pathway. P value was calculated using one-way ANOVA.

    Article Snippet: Primary antibodies are: Ki67 (1:50; Abcam, ab16667), Phospho-Histone H3 (PH3, 1:200; Cell Signaling Technology; 9706L), Aurora B kinase (1:200; BD Transduction Laboratories; 611082), cardiac Troponin T (cTnT, 1:100; Thermo Fisher Scientific, MS-295-P1), cardiac Troponin I (cTnI, 1:200; Abcam; ab47003), Caveolin-1 (1:100, Cell Signaling Technology; 3238S), Wheat Germ Agglutinin (WGA), Alexa Fluor® 633 Conjugate was used on the same sections to outline cardiomyocytes.

    Techniques: Isolation, DNA Synthesis, Microscopy, Immunostaining

    Generation and characterization of cardiac myocyte-specific Hdac3-Tg mice. A , schematic of transgenic construct used to generate Hdac3 -Tg mice. α -MHC , α-myosin heavy chain. B , Hdac3 mRNA expression analysis in two different Hdac3 -Tg mice lines. Transcripts for Hdac3 were detected by qRT-PCR in P1 hearts from wild-type ( WT ) and Hdac3 -Tg mice. Three mice in each group were tested, and values are expressed as the -fold change in transcript abundance (± S.D.) when compared with wild-type mice. C , endogenous and transgenic Hdac3 protein were detected with an Hdac3 antibody. All Western blots were performed at least three times with similar results. D , Hdac3 expression in myocardium of P1 wild-type and Hdac3 -Tg mice was quantified by using ImageJ software. E , increased HDAC activity in Hdac3 -Tg mice. Lysates from P1 hearts were assayed for HDAC activity. Data are the average results (± S.D.) from three separate experiments. F , decreased acetylation of histone H4 in Hdac3 -Tg mice. Western blot analysis of acetylated-histone H4 in myocardium from P1 wild-type and Hdac3 -Tg mice using anti-acetyl-histone H4 antibody was performed. ImageJ software was used to quantify -fold change in acetylation. G , Hdac1 and Hdac2 levels are not changed in Hdac3 -Tg mice. Western blot analysis of myocardium lysates from three P1 wild-type and three Hdac3 -Tg mice using anti-Hdac1 and anti-Hdac2 antibody was performed. α-Tubulin is used as a loading control.

    Journal: The Journal of Biological Chemistry

    Article Title: Transgenic Overexpression of Hdac3 in the Heart Produces Increased Postnatal Cardiac Myocyte Proliferation but Does Not Induce Hypertrophy *

    doi: 10.1074/jbc.M803686200

    Figure Lengend Snippet: Generation and characterization of cardiac myocyte-specific Hdac3-Tg mice. A , schematic of transgenic construct used to generate Hdac3 -Tg mice. α -MHC , α-myosin heavy chain. B , Hdac3 mRNA expression analysis in two different Hdac3 -Tg mice lines. Transcripts for Hdac3 were detected by qRT-PCR in P1 hearts from wild-type ( WT ) and Hdac3 -Tg mice. Three mice in each group were tested, and values are expressed as the -fold change in transcript abundance (± S.D.) when compared with wild-type mice. C , endogenous and transgenic Hdac3 protein were detected with an Hdac3 antibody. All Western blots were performed at least three times with similar results. D , Hdac3 expression in myocardium of P1 wild-type and Hdac3 -Tg mice was quantified by using ImageJ software. E , increased HDAC activity in Hdac3 -Tg mice. Lysates from P1 hearts were assayed for HDAC activity. Data are the average results (± S.D.) from three separate experiments. F , decreased acetylation of histone H4 in Hdac3 -Tg mice. Western blot analysis of acetylated-histone H4 in myocardium from P1 wild-type and Hdac3 -Tg mice using anti-acetyl-histone H4 antibody was performed. ImageJ software was used to quantify -fold change in acetylation. G , Hdac1 and Hdac2 levels are not changed in Hdac3 -Tg mice. Western blot analysis of myocardium lysates from three P1 wild-type and three Hdac3 -Tg mice using anti-Hdac1 and anti-Hdac2 antibody was performed. α-Tubulin is used as a loading control.

    Article Snippet: P1 wild-type and Hdac3 -Tg heart sections were immunostained with anti-phospho-histone H3 antibody (1:2000, Cell Signaling), Ki67 antibody (1:50), and MF-20 (1:25, HybridomaBank).

    Techniques: Mouse Assay, Transgenic Assay, Construct, Expressing, Quantitative RT-PCR, Western Blot, Software, Activity Assay

    Cardiac defects in Hdac3-Tg neonates. A , hematoxylin and  eosin-stained sections at successive levels of P1 hearts from wild-type  ( WT ) and  Hdac3 -Tg mice demonstrate a thicker septum, thicker  ventricular walls, and diminished ventricular lumens.  B , hematoxylin  and eosin-stained sections of adult hearts from wild-type and Hdac3 -Tg mice demonstrate normal ventricular septum, walls, and  lumens.  C , increased proliferation of cardiomyocytes in P1 Hdac3 -Tg mice. Immunohistochemistry for phospho-histone H3 was  performed on heart sections from P1 mice. Quantification of phospho-histone  H3-positive cells was performed on eight sections from three individual hearts  and averaged.  D , co-immunofluorescent staining for Ki67  ( red ), MyHC ( green , MF-20 staining), and  4′,6-diamidino-2-phenylindole ( DAPI ) ( blue ) was  performed on heart sections from P1 wild-type and  Hdac3 -Tg mice. E , wheat germ agglutinin staining shows decreased myocyte size in Hdac3 -Tg when compared with wild-type P1 hearts.

    Journal: The Journal of Biological Chemistry

    Article Title: Transgenic Overexpression of Hdac3 in the Heart Produces Increased Postnatal Cardiac Myocyte Proliferation but Does Not Induce Hypertrophy *

    doi: 10.1074/jbc.M803686200

    Figure Lengend Snippet: Cardiac defects in Hdac3-Tg neonates. A , hematoxylin and eosin-stained sections at successive levels of P1 hearts from wild-type ( WT ) and Hdac3 -Tg mice demonstrate a thicker septum, thicker ventricular walls, and diminished ventricular lumens. B , hematoxylin and eosin-stained sections of adult hearts from wild-type and Hdac3 -Tg mice demonstrate normal ventricular septum, walls, and lumens. C , increased proliferation of cardiomyocytes in P1 Hdac3 -Tg mice. Immunohistochemistry for phospho-histone H3 was performed on heart sections from P1 mice. Quantification of phospho-histone H3-positive cells was performed on eight sections from three individual hearts and averaged. D , co-immunofluorescent staining for Ki67 ( red ), MyHC ( green , MF-20 staining), and 4′,6-diamidino-2-phenylindole ( DAPI ) ( blue ) was performed on heart sections from P1 wild-type and Hdac3 -Tg mice. E , wheat germ agglutinin staining shows decreased myocyte size in Hdac3 -Tg when compared with wild-type P1 hearts.

    Article Snippet: P1 wild-type and Hdac3 -Tg heart sections were immunostained with anti-phospho-histone H3 antibody (1:2000, Cell Signaling), Ki67 antibody (1:50), and MF-20 (1:25, HybridomaBank).

    Techniques: Staining, Mouse Assay, Immunohistochemistry

    Proliferation and initial differentiation of interneuron precursors in the PWM. a–c , Distribution of dividing interneuron precursors in Pax-2-GFP transgenic mice at three different postnatal ages (P2, P6, P9). a , b , Graphs illustrate the frequency of GFP/BrdU double-labeled cells in PWM and cortical layers, 2 h ( a ) or 24 h ( b ) after BrdU administration ( n = 2–4 cases/time point). c–e , The majority of dividing cells are located in the PWM (arrows in c ). This distribution is confirmed by immunostaining for phospho-histone Ph3 ( d , e ). f , Laminar position of GFP/BrdU-double-labeled cells at increasing times after BrdU injection: 4 d after nucleotide analog administration, many cells are still in the PWM. g–n shows Pax-2 immunolabeling ( g–j ) or Pax-2/GFP expression ( k–n ) in interneuron precursors labeled by BrdU 2 h ( g , h , k , l ) or 24 h ( i , j , m , n ) before animals were killed. o , p , Double-labeled cells (arrows) consistently show a weaker Pax-2 expression at 2 h than at 24 h, as confirmed by the quantitative estimation of Pax-2 immunofluorescence intensity ( o , Student's t test, p

    Journal: The Journal of Neuroscience

    Article Title: Laminar Fate and Phenotype Specification of Cerebellar GABAergic Interneurons

    doi: 10.1523/JNEUROSCI.0957-09.2009

    Figure Lengend Snippet: Proliferation and initial differentiation of interneuron precursors in the PWM. a–c , Distribution of dividing interneuron precursors in Pax-2-GFP transgenic mice at three different postnatal ages (P2, P6, P9). a , b , Graphs illustrate the frequency of GFP/BrdU double-labeled cells in PWM and cortical layers, 2 h ( a ) or 24 h ( b ) after BrdU administration ( n = 2–4 cases/time point). c–e , The majority of dividing cells are located in the PWM (arrows in c ). This distribution is confirmed by immunostaining for phospho-histone Ph3 ( d , e ). f , Laminar position of GFP/BrdU-double-labeled cells at increasing times after BrdU injection: 4 d after nucleotide analog administration, many cells are still in the PWM. g–n shows Pax-2 immunolabeling ( g–j ) or Pax-2/GFP expression ( k–n ) in interneuron precursors labeled by BrdU 2 h ( g , h , k , l ) or 24 h ( i , j , m , n ) before animals were killed. o , p , Double-labeled cells (arrows) consistently show a weaker Pax-2 expression at 2 h than at 24 h, as confirmed by the quantitative estimation of Pax-2 immunofluorescence intensity ( o , Student's t test, p

    Article Snippet: Slices obtained from different experimental sets were processed for immunofluorescent labeling with different primary antibodies (all dissolved in PBS with 1.5% normal serum and 0.25% Triton X-100): anti-parvalbumin (1:1500, monoclonal; Swant), Pax-2 (1:200, polyclonal; Zymed), anti-phospho-histone H3 (1:100, polyclonal, Upstate Biotechnology), anti-neurogranin (1:250; polyclonal; Millipore Bioscience Research Reagents), anti-Ng2 (1:200; polyclonal; Millipore Bioscience Research Reagents), anti-S100 (1:1000, monoclonal, Sigma), anti-Olig2 (1:500; polyclonal; Millipore Bioscience Research Reagents), anti-GFP (1:700, polyclonal or monoclonal; Invitrogen), anti-RFP (1:500, Abcam), anti-BrdU (1:500, monoclonal, Sigma) (see for technical details).

    Techniques: Transgenic Assay, Mouse Assay, Labeling, Immunostaining, Injection, Immunolabeling, Expressing, Immunofluorescence

    Overexpression of cyclin B1 reduces the survival of BRCA1-deficient cells. a  Unsynchronized MCF7 cells in which BRCA1 was knocked down and/or cyclin B1 was overexpressed were examined according to the cell cycle phase, determined by flow cytometry. Representative histograms showing the DNA content in MCF7 cells with BRCA1 depletion and/or cyclin B1 overexpression.  b  Percentages of cells in each cell cycle phase are shown.  c  Protein expression patterns in  BRCA1 -knockdown and/or cyclin B1-overexpressing MCF7 cells were analyzed via Western blot analysis.  d  To count the cells in G2/M transition, unsynchronized MCF7 cells transfected with BRCA1 siRNA and/or transduced with Ad-cyclin B1 were stained with phospho (Ser10)-Histone H3 and analyzed via flow cytometry.  e  Survival of cyclin B1-overexpressing MCF7 cells was measured in cells transfected with scrambled siRNA or  BRCA1 -targeted siRNA. The results are expressed as survival relative to that at infection or transfection.  f  A breast cancer cell line derived from a  Brca1 Δ11/Δ11 p53 −/−  mouse was infected with increasing amounts of Ad-cyclin B1, after which survival was analyzed with an MTT assay.  g  A breast cancer cell line derived from a  Brca1 Δ11/Δ11 p53 −/−  mouse was infected with Ad-cyclin B1 or Ad-GFP (control), after which the protein pattern was analyzed via Western blotting.  h  Estimated survival of  Brca1 Δ11/Δ11 ;p53 −/−  mammary tumor cell lines at the indicated concentration of vinblastine.  i  The survival of  Brca1 Δ11/Δ11 p53 −/−  mammary tumor cell lines was assessed after transfection with scramble or  Ccnb1  siRNA and treatment with vinblastine at the indicated concentration. ** P

    Journal: Experimental & Molecular Medicine

    Article Title: Cyclin B1 stability is increased by interaction with BRCA1, and its overexpression suppresses the progression of BRCA1-associated mammary tumors

    doi: 10.1038/s12276-018-0169-z

    Figure Lengend Snippet: Overexpression of cyclin B1 reduces the survival of BRCA1-deficient cells. a Unsynchronized MCF7 cells in which BRCA1 was knocked down and/or cyclin B1 was overexpressed were examined according to the cell cycle phase, determined by flow cytometry. Representative histograms showing the DNA content in MCF7 cells with BRCA1 depletion and/or cyclin B1 overexpression. b Percentages of cells in each cell cycle phase are shown. c Protein expression patterns in BRCA1 -knockdown and/or cyclin B1-overexpressing MCF7 cells were analyzed via Western blot analysis. d To count the cells in G2/M transition, unsynchronized MCF7 cells transfected with BRCA1 siRNA and/or transduced with Ad-cyclin B1 were stained with phospho (Ser10)-Histone H3 and analyzed via flow cytometry. e Survival of cyclin B1-overexpressing MCF7 cells was measured in cells transfected with scrambled siRNA or BRCA1 -targeted siRNA. The results are expressed as survival relative to that at infection or transfection. f A breast cancer cell line derived from a Brca1 Δ11/Δ11 p53 −/− mouse was infected with increasing amounts of Ad-cyclin B1, after which survival was analyzed with an MTT assay. g A breast cancer cell line derived from a Brca1 Δ11/Δ11 p53 −/− mouse was infected with Ad-cyclin B1 or Ad-GFP (control), after which the protein pattern was analyzed via Western blotting. h Estimated survival of Brca1 Δ11/Δ11 ;p53 −/− mammary tumor cell lines at the indicated concentration of vinblastine. i The survival of Brca1 Δ11/Δ11 p53 −/− mammary tumor cell lines was assessed after transfection with scramble or Ccnb1 siRNA and treatment with vinblastine at the indicated concentration. ** P

    Article Snippet: The primary antibodies used were anti-phospho-histone H3 (Merck Millipore) and anti-PCNA (Atlas Antibodies).

    Techniques: Over Expression, Flow Cytometry, Cytometry, Expressing, Western Blot, Transfection, Transduction, Staining, Infection, Derivative Assay, MTT Assay, Concentration Assay

    Lentiviral knockdown of FoxF1, FoxF2 or both in mouse and human RMS cells decreased cellular growth  in vitro (A)  Efficiency of FoxF1 and FoxF2 lentiviral knockdown in mouse 76-9 rhabdomyosarcoma cells was shown by qRT-PCR.  β-actin  mRNA was used for normalization. ( B)  Depletion of FoxF1, FoxF2 or both decreased proliferation of 76-9 cells  in vitro . Control, FoxF1-KD, FoxF2-KD and F1+F2-KD 76-9 cells were seeded in triplicates and counted at different time points using WST1 Cell Proliferation Assay. ( C)  Depletion of FoxF1, FoxF2 or both decreased colony formation in soft agar compared to control RMS cells. Cells were seeded in triplicates. Values are the means ± SD of three independent experiments. A  p value  lt;0.05 is shown with (*). ( D)  Depletion of FoxF1, FoxF2 or both decreased the number of cells in mitosis shown by flow cytometery with phospho-Histone H3 antibodies.  (E)  Efficiency of FoxF1 and FoxF2 lentiviral knockdown in human Rh30 rhabdomyosarcoma cells was shown by qRT-PCR.  β-actin  mRNA was used for normalization. ( F)  Depletion of FoxF1, FoxF2 or both decreased proliferation of Rh30 cells  in vitro . Control, FoxF1-, FoxF2- or F1+F2-deficient Rh30 cells were seeded in triplicates and counted at different time points using hemocytometer. Data represent mean±s.d. of three independent experiments. A  p  value   lt; 0.05 is shown with (*);  p  value   lt; 0.01 is shown with (**).

    Journal: Oncogene

    Article Title: FoxF1 and FoxF2 transcription factors synergistically promote Rhabdomyosarcoma carcinogenesis by repressing transcription of p21Cip1 CDK inhibitor

    doi: 10.1038/onc.2016.254

    Figure Lengend Snippet: Lentiviral knockdown of FoxF1, FoxF2 or both in mouse and human RMS cells decreased cellular growth in vitro (A) Efficiency of FoxF1 and FoxF2 lentiviral knockdown in mouse 76-9 rhabdomyosarcoma cells was shown by qRT-PCR. β-actin mRNA was used for normalization. ( B) Depletion of FoxF1, FoxF2 or both decreased proliferation of 76-9 cells in vitro . Control, FoxF1-KD, FoxF2-KD and F1+F2-KD 76-9 cells were seeded in triplicates and counted at different time points using WST1 Cell Proliferation Assay. ( C) Depletion of FoxF1, FoxF2 or both decreased colony formation in soft agar compared to control RMS cells. Cells were seeded in triplicates. Values are the means ± SD of three independent experiments. A p value lt;0.05 is shown with (*). ( D) Depletion of FoxF1, FoxF2 or both decreased the number of cells in mitosis shown by flow cytometery with phospho-Histone H3 antibodies. (E) Efficiency of FoxF1 and FoxF2 lentiviral knockdown in human Rh30 rhabdomyosarcoma cells was shown by qRT-PCR. β-actin mRNA was used for normalization. ( F) Depletion of FoxF1, FoxF2 or both decreased proliferation of Rh30 cells in vitro . Control, FoxF1-, FoxF2- or F1+F2-deficient Rh30 cells were seeded in triplicates and counted at different time points using hemocytometer. Data represent mean±s.d. of three independent experiments. A p value lt; 0.05 is shown with (*); p value lt; 0.01 is shown with (**).

    Article Snippet: Immunostaining was performed using following antibodies: anti-myogenin, anti-Ki67, anti-phospho-Histone H3 (Santa Cruz), anti-pRb, anti-p27Kip1 , anti-p21Cip1 (Cell Signaling Technology, Inc.), anti-Cyclin D1 (BD biosciences) were done as previously described ( , ).

    Techniques: In Vitro, Quantitative RT-PCR, Proliferation Assay, Flow Cytometry

    SHP-2 N308D leads to a delay of the cardiac cell cycle. ( A-D ) Transverse heart sections through stage 33 (A,B) and stage 37 (C,D) Xenopus embryos stained with Tmy (green) and anti-phospho histone H3 (red), from uninjected (A,C) and SHP-2 N308D -derived (B,D)

    Journal: Development (Cambridge, England)

    Article Title: SHP-2 acts via ROCK to regulate the cardiac actin cytoskeleton

    doi: 10.1242/dev.067579

    Figure Lengend Snippet: SHP-2 N308D leads to a delay of the cardiac cell cycle. ( A-D ) Transverse heart sections through stage 33 (A,B) and stage 37 (C,D) Xenopus embryos stained with Tmy (green) and anti-phospho histone H3 (red), from uninjected (A,C) and SHP-2 N308D -derived (B,D)

    Article Snippet: Antibodies used in immunohistochemistry were: mouse anti-tropomyosin (1:50), mouse anti-troponin (1:20), mouse anti-fibrillin (1:50) (all from Developmental Studies Hybridoma Bank), mouse anti-MHC (1:500; Abcam), rabbit anti-fibronectin (1:50; Sigma), rabbit anti-phospho histone H3 (1:50; Upstate) and rabbit anti-cleaved caspase-3 (1:50; Cell Signaling).

    Techniques: Staining, Derivative Assay