rabbit anti phospho histone 3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho histone 3 ser10
    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for <t>phospho-histone-3</t> (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Rabbit Anti Phospho Histone 3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "GU81, a VEGFR2 antagonist peptoid, enhances the anti-tumor activity of doxorubicin in the murine MMTV-PyMT transgenic model of breast cancer"

    Article Title: GU81, a VEGFR2 antagonist peptoid, enhances the anti-tumor activity of doxorubicin in the murine MMTV-PyMT transgenic model of breast cancer

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-397

    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Figure Legend Snippet: Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.

    Techniques Used: Immunohistochemistry, Software, MANN-WHITNEY

    rabbit anti phospho histone 3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho histone 3 ser10
    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for <t>phospho-histone-3</t> (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Rabbit Anti Phospho Histone 3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "GU81, a VEGFR2 antagonist peptoid, enhances the anti-tumor activity of doxorubicin in the murine MMTV-PyMT transgenic model of breast cancer"

    Article Title: GU81, a VEGFR2 antagonist peptoid, enhances the anti-tumor activity of doxorubicin in the murine MMTV-PyMT transgenic model of breast cancer

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-397

    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Figure Legend Snippet: Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.

    Techniques Used: Immunohistochemistry, Software, MANN-WHITNEY

    phospho histone h3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone h3 ser10
    Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit monoclonal anti phospho histone h3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti phospho histone h3 ser10
    PGE 2 -dependent effect on phospho-histone H3 <t>(Ser10)</t> expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.
    Rabbit Monoclonal Anti Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti phospho histone h3 ser10/product/Cell Signaling Technology Inc
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    1) Product Images from "Prostaglandin E2 alters Wnt-dependent migration and proliferation in neuroectodermal stem cells: implications for autism spectrum disorders"

    Article Title: Prostaglandin E2 alters Wnt-dependent migration and proliferation in neuroectodermal stem cells: implications for autism spectrum disorders

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/1478-811X-12-19

    PGE 2 -dependent effect on phospho-histone H3 (Ser10) expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.
    Figure Legend Snippet: PGE 2 -dependent effect on phospho-histone H3 (Ser10) expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.

    Techniques Used: Expressing, Western Blot

    phospho histone h3 ser10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone h3 ser10
    Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho histone h3 ser10/product/Cell Signaling Technology Inc
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    phospho h3 serine 10  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho h3 serine 10
    Summary of ERRβ splice variant dominance and resulting phenotype
    Phospho H3 Serine 10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "ERRβ splice variants differentially regulate cell cycle progression"

    Article Title: ERRβ splice variants differentially regulate cell cycle progression

    Journal: Cell Cycle

    doi: 10.4161/15384101.2014.972886

    Summary of ERRβ splice variant dominance and resulting phenotype
    Figure Legend Snippet: Summary of ERRβ splice variant dominance and resulting phenotype

    Techniques Used: Variant Assay

    rabbit monoclonal anti phospho histone h3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti phospho histone h3
    (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone <t>H3(Ser10)</t> in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.
    Rabbit Monoclonal Anti Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Evolution of Resistance to Aurora Kinase B Inhibitors in Leukaemia Cells"

    Article Title: Evolution of Resistance to Aurora Kinase B Inhibitors in Leukaemia Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030734

    (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone H3(Ser10) in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.
    Figure Legend Snippet: (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone H3(Ser10) in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    histone h3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc histone h3
    Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone <t>H3</t> labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.
    Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Tumor Environmental Factors Glucose Deprivation and Lactic Acidosis Induce Mitotic Chromosomal Instability – An Implication in Aneuploid Human Tumors"

    Article Title: Tumor Environmental Factors Glucose Deprivation and Lactic Acidosis Induce Mitotic Chromosomal Instability – An Implication in Aneuploid Human Tumors

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0063054

    Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone H3 labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.
    Figure Legend Snippet: Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone H3 labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.

    Techniques Used: Cell Culture

    phospho histone h3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho histone h3
    (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone <t>H3</t> (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.
    Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina"

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0053517

    (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone H3 (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.
    Figure Legend Snippet: (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone H3 (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.

    Techniques Used: Immunofluorescence, Staining, Labeling

    (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY294002, fixed and labeled with DAPI (blue) or antiserum against phospho-histone H3 (P-H3) (red). Control explants were cultured for the same period without inhibitor. Note the increase in the number of p-H3-positive cells at the border of the treated retina. (B) Explants of retinas from embryo at E7 were treated with 25 µM LY 294002 for 12 or 24 h and processed for P-H3 detection by western blot. (C) Retinal monolayer cultures at E7C1 were treated for 24 h with 10 µM or 25 µM LY 294002 and processed for P-H3 detection. Protein gel loading was assessed with anti-α-tubulin and anti-ERK antisera, respectively. Representative blots from at least 3 independent experiments are shown. (D) Quantification of blots represented in B and C. Data represent the mean ± S.E.M. in arbitrary units (A.U.) of 5–6 experiments performed in duplicate or triplicate. **p<0.01 and *p<0.05, compared to control cultures without LY treatment. Ct = control cultures or explants cultivated without drugs. NBL = Neuroblastic Layer. GCL = Prospective Ganglion Cell Layer. Scale bar = 20 µm.
    Figure Legend Snippet: (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY294002, fixed and labeled with DAPI (blue) or antiserum against phospho-histone H3 (P-H3) (red). Control explants were cultured for the same period without inhibitor. Note the increase in the number of p-H3-positive cells at the border of the treated retina. (B) Explants of retinas from embryo at E7 were treated with 25 µM LY 294002 for 12 or 24 h and processed for P-H3 detection by western blot. (C) Retinal monolayer cultures at E7C1 were treated for 24 h with 10 µM or 25 µM LY 294002 and processed for P-H3 detection. Protein gel loading was assessed with anti-α-tubulin and anti-ERK antisera, respectively. Representative blots from at least 3 independent experiments are shown. (D) Quantification of blots represented in B and C. Data represent the mean ± S.E.M. in arbitrary units (A.U.) of 5–6 experiments performed in duplicate or triplicate. **p<0.01 and *p<0.05, compared to control cultures without LY treatment. Ct = control cultures or explants cultivated without drugs. NBL = Neuroblastic Layer. GCL = Prospective Ganglion Cell Layer. Scale bar = 20 µm.

    Techniques Used: Labeling, Cell Culture, Western Blot

    (A–D) Retinal cell cultures at E7C1 were fixed and assayed by immunocytochemistry for phospho-4E-BP1 (A) and phospho-histone H3 (B). DNA was stained with DAPI (C) and merged figures are shown in (D). Arrows point to double labeled cells. (E-G) Phospho-4E-BP1 labeling in mitotic cells of 8-day-old chick embryo retinas. Retinal sections were stained with anti-phospho-4E-BP1 (E) and anti-phospho-histone H3 (F). Merged figures are shown in (G). Phospho-4E-BP1 labeled cells were confined to the ventricular margin of the retina. (H) Detection of phospho-4E-BP1 in extracts of retinal cultures at E7C1 without any treatment. The phosphorylated forms γ and β of the protein are indicated by arrows, respectively. (I) Expression of phospho-4E-BP1 in extracts of retinal explants treated with 25 µM LY 294002 for 4, 8 or 12 h. Representative blots from at least 3 independent experiments are shown. Ct = control explants cultivated without drug. NBL = Neuroblastic Layer. Scale bar = 20 µm.
    Figure Legend Snippet: (A–D) Retinal cell cultures at E7C1 were fixed and assayed by immunocytochemistry for phospho-4E-BP1 (A) and phospho-histone H3 (B). DNA was stained with DAPI (C) and merged figures are shown in (D). Arrows point to double labeled cells. (E-G) Phospho-4E-BP1 labeling in mitotic cells of 8-day-old chick embryo retinas. Retinal sections were stained with anti-phospho-4E-BP1 (E) and anti-phospho-histone H3 (F). Merged figures are shown in (G). Phospho-4E-BP1 labeled cells were confined to the ventricular margin of the retina. (H) Detection of phospho-4E-BP1 in extracts of retinal cultures at E7C1 without any treatment. The phosphorylated forms γ and β of the protein are indicated by arrows, respectively. (I) Expression of phospho-4E-BP1 in extracts of retinal explants treated with 25 µM LY 294002 for 4, 8 or 12 h. Representative blots from at least 3 independent experiments are shown. Ct = control explants cultivated without drug. NBL = Neuroblastic Layer. Scale bar = 20 µm.

    Techniques Used: Immunocytochemistry, Staining, Labeling, Expressing

    (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY 294002, fixed and sections processed by TUNEL assays as described in methods. Representative micrographs of TUNEL labeling (green) are shown. DAPI (blue) was used to label nuclei of all cells and explants were oriented with their outer, ventricular margin at the upper side of the micrographs. Data are representative of three experiments. (B) Retinal explants were treated for 22 h with 25 µM LY 294002, fixed and sections processed for immunocytochemistry against cleaved caspase-3 (green) and phospho-histone H3 (red). (C) Quantification of phospho-histone H3 and cleaved caspase 3 positive cells in retinal explants. Positive cells were counted in 10 transverse sections of retinal explants with 150 µm of linear extent parallel to their surface that were divided in 3 segments with the same width (o = outer segment; m = medium segment; I = inner segment). Data represent the mean ± S.E.M. of cell counts/segment from four independent experiments. Scale bar = 20 µm.
    Figure Legend Snippet: (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY 294002, fixed and sections processed by TUNEL assays as described in methods. Representative micrographs of TUNEL labeling (green) are shown. DAPI (blue) was used to label nuclei of all cells and explants were oriented with their outer, ventricular margin at the upper side of the micrographs. Data are representative of three experiments. (B) Retinal explants were treated for 22 h with 25 µM LY 294002, fixed and sections processed for immunocytochemistry against cleaved caspase-3 (green) and phospho-histone H3 (red). (C) Quantification of phospho-histone H3 and cleaved caspase 3 positive cells in retinal explants. Positive cells were counted in 10 transverse sections of retinal explants with 150 µm of linear extent parallel to their surface that were divided in 3 segments with the same width (o = outer segment; m = medium segment; I = inner segment). Data represent the mean ± S.E.M. of cell counts/segment from four independent experiments. Scale bar = 20 µm.

    Techniques Used: TUNEL Assay, Labeling, Immunocytochemistry

    (A) Monolayer cultures of retinal cells from E7 were established as described. After 2 h, LY294002 to a final concentration of 25 µM was added. After 24 h, medium was removed, fresh medium was added and cells cultivated for an additional 24 h period. At the end of incubations, protein extracts were analyzed for the indicated proteins by western blotting. Representative blots are shown. (B) Blots were quantified by densitometry and data are expressed as the mean ± S.E.M. (% control) of 3 or 4 experiments. ***p<0.001 relative to control without treatment. ## p<0.01 relative to cultures treated on the second day or during the entire period. (C) Expression of transitin in treated explants. Retinal explants from 7-day-old chick embryos were treated for 24 h with 25 µM LY294002, fixed and labeled with anti-pH3 (green) or antiserum against transitin (red). Arrows point to double labeled cells. Control explants were cultured for the same period without inhibitor. Scale bar = 10 µm.
    Figure Legend Snippet: (A) Monolayer cultures of retinal cells from E7 were established as described. After 2 h, LY294002 to a final concentration of 25 µM was added. After 24 h, medium was removed, fresh medium was added and cells cultivated for an additional 24 h period. At the end of incubations, protein extracts were analyzed for the indicated proteins by western blotting. Representative blots are shown. (B) Blots were quantified by densitometry and data are expressed as the mean ± S.E.M. (% control) of 3 or 4 experiments. ***p<0.001 relative to control without treatment. ## p<0.01 relative to cultures treated on the second day or during the entire period. (C) Expression of transitin in treated explants. Retinal explants from 7-day-old chick embryos were treated for 24 h with 25 µM LY294002, fixed and labeled with anti-pH3 (green) or antiserum against transitin (red). Arrows point to double labeled cells. Control explants were cultured for the same period without inhibitor. Scale bar = 10 µm.

    Techniques Used: Concentration Assay, Western Blot, Expressing, Labeling, Cell Culture

    phosphor ser10 histone h3  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc phosphor ser10 histone h3
    Phosphor Ser10 Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho ser10 histone h3  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho ser10 histone h3
    (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for <t>phospho(Ser10)-histone</t> H3 (p-H3) were measured for crude cell extracts as described for part A.
    Phospho Ser10 Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Pharmacological AMP Kinase Activators Target the Nucleolar Organization and Control Cell Proliferation"

    Article Title: Pharmacological AMP Kinase Activators Target the Nucleolar Organization and Control Cell Proliferation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0088087

    (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for phospho(Ser10)-histone H3 (p-H3) were measured for crude cell extracts as described for part A.
    Figure Legend Snippet: (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for phospho(Ser10)-histone H3 (p-H3) were measured for crude cell extracts as described for part A.

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    Cell Signaling Technology Inc rabbit anti phospho histone 3 ser10
    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for <t>phospho-histone-3</t> (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Rabbit Anti Phospho Histone 3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho histone h3 ser10
    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for <t>phospho-histone-3</t> (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.
    Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit monoclonal anti phospho histone h3 ser10
    PGE 2 -dependent effect on phospho-histone H3 <t>(Ser10)</t> expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.
    Rabbit Monoclonal Anti Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho h3 serine 10
    Summary of ERRβ splice variant dominance and resulting phenotype
    Phospho H3 Serine 10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit monoclonal anti phospho histone h3
    (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone <t>H3(Ser10)</t> in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.
    Rabbit Monoclonal Anti Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc histone h3
    Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone <t>H3</t> labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.
    Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho histone h3
    (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone <t>H3</t> (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.
    Phospho Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphor ser10 histone h3
    (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone <t>H3</t> (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.
    Phosphor Ser10 Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho ser10 histone h3
    (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for <t>phospho(Ser10)-histone</t> H3 (p-H3) were measured for crude cell extracts as described for part A.
    Phospho Ser10 Histone H3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.

    Journal: BMC Cancer

    Article Title: GU81, a VEGFR2 antagonist peptoid, enhances the anti-tumor activity of doxorubicin in the murine MMTV-PyMT transgenic model of breast cancer

    doi: 10.1186/1471-2407-10-397

    Figure Lengend Snippet: Combination therapy of doxorubicin and GU81 reduces the growth index of MMTV-PyMT tumors . Tumor sections from each treatment group were evaluated for phospho-histone-3 (A&B) and active-caspase-3 (C&D) by immunohistochemistry as described in the methods section. Signal intensity was quantified using Elements software and is displayed and mean +/- SEM. All quantification includes 3 animals/group and 5 sections/animal. D) A growth index was calculated whereby the number of phospho-histone-3 positive cells (actively proliferating) was divided by the number of active caspase-3 positive cells (undergoing apoptosis). *, p < 0.05; **, p < 0.01 vs control, 1-way ANOVA and Mann Whitney test.

    Article Snippet: Primary antibodies were used at a final concentration of 5-10 μg/ml and include: rabbit anti-perilipin [ ], rabbit anti-adiponectin [ ], rabbit anti-phospho-histone-3 (Ser10) (Upstate, Lake Placid, NY), rabbit anti-cleaved caspase-3 (Asp 175) (Cell Signaling), rat anti-endomucin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), chicken anti-VEGF (Abcam, Cambridge, MA), and goat anti-F4/80 (Santa Cruz Biotechnology).

    Techniques: Immunohistochemistry, Software, MANN-WHITNEY

    PGE 2 -dependent effect on phospho-histone H3 (Ser10) expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.

    Journal: Cell Communication and Signaling : CCS

    Article Title: Prostaglandin E2 alters Wnt-dependent migration and proliferation in neuroectodermal stem cells: implications for autism spectrum disorders

    doi: 10.1186/1478-811X-12-19

    Figure Lengend Snippet: PGE 2 -dependent effect on phospho-histone H3 (Ser10) expression. Western blot analysis was used to determine Phospho-Histone H3 (Ser10) protein (17 kDa). The expression of Phospho-Histone H3 (Ser10) represented in fold change was 1, 1.04, 1.35, 1.52, 1.36, and 1.58, respectively. The error bars represent + SEM and values were considered significantly different from untreated * p < 0.05, ** p < 0.01. Average measurements represent protein from three independent experiments ( N = 3). β-Actin was used to indicate equal loading.

    Article Snippet: Detection of rabbit monoclonal anti-Phospho-Histone H3 (Ser10) (1:1000; Cell Signaling) was used as a measure of cell splitting behaviour.

    Techniques: Expressing, Western Blot

    Summary of ERRβ splice variant dominance and resulting phenotype

    Journal: Cell Cycle

    Article Title: ERRβ splice variants differentially regulate cell cycle progression

    doi: 10.4161/15384101.2014.972886

    Figure Lengend Snippet: Summary of ERRβ splice variant dominance and resulting phenotype

    Article Snippet: Membranes were blocked in 5% nonfat dry milk buffer, unless otherwise noted, and incubated overnight at 4°C with primary antibodies for: PARP (1:1000, http://www.cellsignal.com/products/primary-antibodies/9542 ), phospho-H3 serine 10 (1:1000, http://www.cellsignal.com/product/productDetail.jsp?productId = 3377) (all from Cell Signaling), p53 (1:1000, Millipore, http://www.millipore.com/catalog/item/05-224 ) p21 (1:300, Santa Cruz Biotechnology, http://www.scbt.com/datasheet-756-p21-h-164-antibody.html ), ERRβ (1:1000, clone H6707 (cl.07) http://www.rndsystems.com/Products/PP-H6707-00 ) and 1:500 clone H6705 (cl.05) http://www.rndsystems.com/Products/PP-H6705-00 ), R&D Systems manufactured by Perseus Proteomics), ERRγ (1:100, Abcam, http://www.abcam.com/estrogen-related-receptor-gamma-antibody-ab82319.html ).

    Techniques: Variant Assay

    (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone H3(Ser10) in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.

    Journal: PLoS ONE

    Article Title: Evolution of Resistance to Aurora Kinase B Inhibitors in Leukaemia Cells

    doi: 10.1371/journal.pone.0030734

    Figure Lengend Snippet: (A) AurkB gene expression as determined by real-time PCR. Expression is displayed as relative ΔΔCt values of CEM/AKB4 compared to that for CEM with Ct values normalised to the cyclophilin-A gene (PPIA). (B) Aurora B protein expression determined by western blot. The densitometric volume of the Aurora B band is expressed relative to the densitometric volume of the loading control gene GAPDH. Error bars represent the SEM of three independent experiments. * p<0.05, ** p<0.005. (C) Detection of phospho Histone H3(Ser10) in CEM and CEM/AKB4 cells treated for 24 hr with increasing concentrations of ZM447439 by western blotting. Shown are representative blots from three independent experiments.

    Article Snippet: Primary antibodies used were rabbit monoclonal anti-Aurora kinase B ([EP1009Y], Abcam), rabbit monoclonal anti-phospho Histone H3(Ser10) ([D2C8], Cell Signaling), rabbit anti-cleaved PARP (Asp214) (Cell Signaling) and mouse monoclonal anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) ([6c5], Abcam).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone H3 labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.

    Journal: PLoS ONE

    Article Title: Tumor Environmental Factors Glucose Deprivation and Lactic Acidosis Induce Mitotic Chromosomal Instability – An Implication in Aneuploid Human Tumors

    doi: 10.1371/journal.pone.0063054

    Figure Lengend Snippet: Cells were cultured in RPMI-1640 medium containing 0.5 mM glucose with lactic acidosis for 7 days. The cells surviving through glucose deprivation with lactic acidosis were then cultured in fresh medium. (A) & (B) Cells were collected at indicated time for analysis of cell cycle and phospho-Histone H3 labelling analysis. (C) The cell growth after release from stress upon nutrition restoration. The results were confirmed by 2 independent experiments.

    Article Snippet: After fixation, the cells were washed with PBS, and permeabilized with 0.3% Triton X-100 in PBS for 15 min. Then the cells were blocked in 1% BSA for 10 min and stained with antibody that specifically recognizes the phosphorylated form of histone H3 (1∶1000 dilution, 3377s, Cell signalling, USA).

    Techniques: Cell Culture

    (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone H3 (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.

    Journal: PLoS ONE

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    doi: 10.1371/journal.pone.0053517

    Figure Lengend Snippet: (A–D) Retinal cell cultures from 7-day-old chick embryos maintained for 1 day (E7C1) were fixed and assayed for immunofluorescence against phospho-Akt (A) and phospho-histone H3 (B). Nuclei were stained with DAPI (C). Merged figures in D. (E–H) Retinal sections from 8-day-old embryo retinas were assayed for immunofluorescence against phospho-Akt (E) and phospho-histone H3 (F). DAPI staining of nuclei (G). Merged figures in (H). Arrows point to double labeled mitotic cells. Scale bar = 20 µm in A–D and 30 µm in E–H.

    Article Snippet: Antibodies against phospho-Akt (Ser473, (catalog # 4060), Akt (catalog # 4691), phospho-4E-BP1 (Thr37/46, catalog # 2855), phospho-Histone H3 (Ser10, catalog # 3377 and # 9706), phospho-GSK-3 (Ser21/9, catalog # 9331), cyclin B1 (catalog # 4138), phospho-CDK1 (Tyr15, catalog # 9111), CDK1 (catalog # 9112), PCNA (catalog # 2586), ERK (catalog # 9102), cleaved caspase-3 (catalog # 9664) were from Cell Signaling Technology (MA, USA).

    Techniques: Immunofluorescence, Staining, Labeling

    (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY294002, fixed and labeled with DAPI (blue) or antiserum against phospho-histone H3 (P-H3) (red). Control explants were cultured for the same period without inhibitor. Note the increase in the number of p-H3-positive cells at the border of the treated retina. (B) Explants of retinas from embryo at E7 were treated with 25 µM LY 294002 for 12 or 24 h and processed for P-H3 detection by western blot. (C) Retinal monolayer cultures at E7C1 were treated for 24 h with 10 µM or 25 µM LY 294002 and processed for P-H3 detection. Protein gel loading was assessed with anti-α-tubulin and anti-ERK antisera, respectively. Representative blots from at least 3 independent experiments are shown. (D) Quantification of blots represented in B and C. Data represent the mean ± S.E.M. in arbitrary units (A.U.) of 5–6 experiments performed in duplicate or triplicate. **p<0.01 and *p<0.05, compared to control cultures without LY treatment. Ct = control cultures or explants cultivated without drugs. NBL = Neuroblastic Layer. GCL = Prospective Ganglion Cell Layer. Scale bar = 20 µm.

    Journal: PLoS ONE

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    doi: 10.1371/journal.pone.0053517

    Figure Lengend Snippet: (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY294002, fixed and labeled with DAPI (blue) or antiserum against phospho-histone H3 (P-H3) (red). Control explants were cultured for the same period without inhibitor. Note the increase in the number of p-H3-positive cells at the border of the treated retina. (B) Explants of retinas from embryo at E7 were treated with 25 µM LY 294002 for 12 or 24 h and processed for P-H3 detection by western blot. (C) Retinal monolayer cultures at E7C1 were treated for 24 h with 10 µM or 25 µM LY 294002 and processed for P-H3 detection. Protein gel loading was assessed with anti-α-tubulin and anti-ERK antisera, respectively. Representative blots from at least 3 independent experiments are shown. (D) Quantification of blots represented in B and C. Data represent the mean ± S.E.M. in arbitrary units (A.U.) of 5–6 experiments performed in duplicate or triplicate. **p<0.01 and *p<0.05, compared to control cultures without LY treatment. Ct = control cultures or explants cultivated without drugs. NBL = Neuroblastic Layer. GCL = Prospective Ganglion Cell Layer. Scale bar = 20 µm.

    Article Snippet: Antibodies against phospho-Akt (Ser473, (catalog # 4060), Akt (catalog # 4691), phospho-4E-BP1 (Thr37/46, catalog # 2855), phospho-Histone H3 (Ser10, catalog # 3377 and # 9706), phospho-GSK-3 (Ser21/9, catalog # 9331), cyclin B1 (catalog # 4138), phospho-CDK1 (Tyr15, catalog # 9111), CDK1 (catalog # 9112), PCNA (catalog # 2586), ERK (catalog # 9102), cleaved caspase-3 (catalog # 9664) were from Cell Signaling Technology (MA, USA).

    Techniques: Labeling, Cell Culture, Western Blot

    (A–D) Retinal cell cultures at E7C1 were fixed and assayed by immunocytochemistry for phospho-4E-BP1 (A) and phospho-histone H3 (B). DNA was stained with DAPI (C) and merged figures are shown in (D). Arrows point to double labeled cells. (E-G) Phospho-4E-BP1 labeling in mitotic cells of 8-day-old chick embryo retinas. Retinal sections were stained with anti-phospho-4E-BP1 (E) and anti-phospho-histone H3 (F). Merged figures are shown in (G). Phospho-4E-BP1 labeled cells were confined to the ventricular margin of the retina. (H) Detection of phospho-4E-BP1 in extracts of retinal cultures at E7C1 without any treatment. The phosphorylated forms γ and β of the protein are indicated by arrows, respectively. (I) Expression of phospho-4E-BP1 in extracts of retinal explants treated with 25 µM LY 294002 for 4, 8 or 12 h. Representative blots from at least 3 independent experiments are shown. Ct = control explants cultivated without drug. NBL = Neuroblastic Layer. Scale bar = 20 µm.

    Journal: PLoS ONE

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    doi: 10.1371/journal.pone.0053517

    Figure Lengend Snippet: (A–D) Retinal cell cultures at E7C1 were fixed and assayed by immunocytochemistry for phospho-4E-BP1 (A) and phospho-histone H3 (B). DNA was stained with DAPI (C) and merged figures are shown in (D). Arrows point to double labeled cells. (E-G) Phospho-4E-BP1 labeling in mitotic cells of 8-day-old chick embryo retinas. Retinal sections were stained with anti-phospho-4E-BP1 (E) and anti-phospho-histone H3 (F). Merged figures are shown in (G). Phospho-4E-BP1 labeled cells were confined to the ventricular margin of the retina. (H) Detection of phospho-4E-BP1 in extracts of retinal cultures at E7C1 without any treatment. The phosphorylated forms γ and β of the protein are indicated by arrows, respectively. (I) Expression of phospho-4E-BP1 in extracts of retinal explants treated with 25 µM LY 294002 for 4, 8 or 12 h. Representative blots from at least 3 independent experiments are shown. Ct = control explants cultivated without drug. NBL = Neuroblastic Layer. Scale bar = 20 µm.

    Article Snippet: Antibodies against phospho-Akt (Ser473, (catalog # 4060), Akt (catalog # 4691), phospho-4E-BP1 (Thr37/46, catalog # 2855), phospho-Histone H3 (Ser10, catalog # 3377 and # 9706), phospho-GSK-3 (Ser21/9, catalog # 9331), cyclin B1 (catalog # 4138), phospho-CDK1 (Tyr15, catalog # 9111), CDK1 (catalog # 9112), PCNA (catalog # 2586), ERK (catalog # 9102), cleaved caspase-3 (catalog # 9664) were from Cell Signaling Technology (MA, USA).

    Techniques: Immunocytochemistry, Staining, Labeling, Expressing

    (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY 294002, fixed and sections processed by TUNEL assays as described in methods. Representative micrographs of TUNEL labeling (green) are shown. DAPI (blue) was used to label nuclei of all cells and explants were oriented with their outer, ventricular margin at the upper side of the micrographs. Data are representative of three experiments. (B) Retinal explants were treated for 22 h with 25 µM LY 294002, fixed and sections processed for immunocytochemistry against cleaved caspase-3 (green) and phospho-histone H3 (red). (C) Quantification of phospho-histone H3 and cleaved caspase 3 positive cells in retinal explants. Positive cells were counted in 10 transverse sections of retinal explants with 150 µm of linear extent parallel to their surface that were divided in 3 segments with the same width (o = outer segment; m = medium segment; I = inner segment). Data represent the mean ± S.E.M. of cell counts/segment from four independent experiments. Scale bar = 20 µm.

    Journal: PLoS ONE

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    doi: 10.1371/journal.pone.0053517

    Figure Lengend Snippet: (A) Retinal explants from 7-day-old chick embryos were treated for 22 h with 25 µM LY 294002, fixed and sections processed by TUNEL assays as described in methods. Representative micrographs of TUNEL labeling (green) are shown. DAPI (blue) was used to label nuclei of all cells and explants were oriented with their outer, ventricular margin at the upper side of the micrographs. Data are representative of three experiments. (B) Retinal explants were treated for 22 h with 25 µM LY 294002, fixed and sections processed for immunocytochemistry against cleaved caspase-3 (green) and phospho-histone H3 (red). (C) Quantification of phospho-histone H3 and cleaved caspase 3 positive cells in retinal explants. Positive cells were counted in 10 transverse sections of retinal explants with 150 µm of linear extent parallel to their surface that were divided in 3 segments with the same width (o = outer segment; m = medium segment; I = inner segment). Data represent the mean ± S.E.M. of cell counts/segment from four independent experiments. Scale bar = 20 µm.

    Article Snippet: Antibodies against phospho-Akt (Ser473, (catalog # 4060), Akt (catalog # 4691), phospho-4E-BP1 (Thr37/46, catalog # 2855), phospho-Histone H3 (Ser10, catalog # 3377 and # 9706), phospho-GSK-3 (Ser21/9, catalog # 9331), cyclin B1 (catalog # 4138), phospho-CDK1 (Tyr15, catalog # 9111), CDK1 (catalog # 9112), PCNA (catalog # 2586), ERK (catalog # 9102), cleaved caspase-3 (catalog # 9664) were from Cell Signaling Technology (MA, USA).

    Techniques: TUNEL Assay, Labeling, Immunocytochemistry

    (A) Monolayer cultures of retinal cells from E7 were established as described. After 2 h, LY294002 to a final concentration of 25 µM was added. After 24 h, medium was removed, fresh medium was added and cells cultivated for an additional 24 h period. At the end of incubations, protein extracts were analyzed for the indicated proteins by western blotting. Representative blots are shown. (B) Blots were quantified by densitometry and data are expressed as the mean ± S.E.M. (% control) of 3 or 4 experiments. ***p<0.001 relative to control without treatment. ## p<0.01 relative to cultures treated on the second day or during the entire period. (C) Expression of transitin in treated explants. Retinal explants from 7-day-old chick embryos were treated for 24 h with 25 µM LY294002, fixed and labeled with anti-pH3 (green) or antiserum against transitin (red). Arrows point to double labeled cells. Control explants were cultured for the same period without inhibitor. Scale bar = 10 µm.

    Journal: PLoS ONE

    Article Title: Inhibition of PI3K/Akt Pathway Impairs G2/M Transition of Cell Cycle in Late Developing Progenitors of the Avian Embryo Retina

    doi: 10.1371/journal.pone.0053517

    Figure Lengend Snippet: (A) Monolayer cultures of retinal cells from E7 were established as described. After 2 h, LY294002 to a final concentration of 25 µM was added. After 24 h, medium was removed, fresh medium was added and cells cultivated for an additional 24 h period. At the end of incubations, protein extracts were analyzed for the indicated proteins by western blotting. Representative blots are shown. (B) Blots were quantified by densitometry and data are expressed as the mean ± S.E.M. (% control) of 3 or 4 experiments. ***p<0.001 relative to control without treatment. ## p<0.01 relative to cultures treated on the second day or during the entire period. (C) Expression of transitin in treated explants. Retinal explants from 7-day-old chick embryos were treated for 24 h with 25 µM LY294002, fixed and labeled with anti-pH3 (green) or antiserum against transitin (red). Arrows point to double labeled cells. Control explants were cultured for the same period without inhibitor. Scale bar = 10 µm.

    Article Snippet: Antibodies against phospho-Akt (Ser473, (catalog # 4060), Akt (catalog # 4691), phospho-4E-BP1 (Thr37/46, catalog # 2855), phospho-Histone H3 (Ser10, catalog # 3377 and # 9706), phospho-GSK-3 (Ser21/9, catalog # 9331), cyclin B1 (catalog # 4138), phospho-CDK1 (Tyr15, catalog # 9111), CDK1 (catalog # 9112), PCNA (catalog # 2586), ERK (catalog # 9102), cleaved caspase-3 (catalog # 9664) were from Cell Signaling Technology (MA, USA).

    Techniques: Concentration Assay, Western Blot, Expressing, Labeling, Cell Culture

    (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for phospho(Ser10)-histone H3 (p-H3) were measured for crude cell extracts as described for part A.

    Journal: PLoS ONE

    Article Title: Pharmacological AMP Kinase Activators Target the Nucleolar Organization and Control Cell Proliferation

    doi: 10.1371/journal.pone.0088087

    Figure Lengend Snippet: (A) Kidney cells were treated with vehicle or the pharmacological compound indicated. Crude extracts were probed with antibodies against B23, fibrillarin, nucleolin or RPA194; actin provided a loading control. The abundance was calculated as nucleolar protein/actin for at least three independent experiments; results were defined as 1 for control samples. Data are shown as averages+SEM; significant differences are marked with * p <0.05 or ** p <0.01. (B) Signals for phospho(Ser10)-histone H3 (p-H3) were measured for crude cell extracts as described for part A.

    Article Snippet: In addition, antibodies against nucleolin (sc-55486; 1∶500), phospho(Ser10)-histone H3 (Cell Signaling, #3377; 1∶1,000), cleaved lamin A (Cell Signaling, #2031; 1∶500); lactate dehydrogenase (Rockland; 1∶4,000), PARP1 (sc-25780; 1∶1,000) or actin (Chemicon; 1∶100,000) were diluted as indicated.

    Techniques: