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  • 99
    Millipore anti brdu antibody
    GH treatment promotes the proliferation of <t>SGZ</t> neurospheres. A) Neurospheres growing in defined media were treated for 24 h with GH (500 ng/mL), pegvisomant (Peg, 20 μg/mL), or GH + pegvisomant. Control cells were treated with saline. Four hours before the end of the treatment period cells were given a <t>BrdU</t> pulse (10 μM). Neurospheres were then dissociated, cells were collected by centrifugation onto coated cover slips, and BrdU was detected by immunocytochemistry. Each bar represents the mean + SEM of 3 experiments in triplicate. * = p
    Anti Brdu Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1229 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend anti brdu ab
    TLR9 expression affected the cell cycle but not apoptosis. ( a ) Caski cells stably transduced with GFP (left panel) or TLR9 (right panel) encoded pbabe were stained with Annexin V and propidium iodide to determine percentage of apoptotic and necrotic cells. ( b ) HNSCC 136 cells were stably transduced with pbabe (left panel), pbabe -GFP (middle panel) or pbabe-TLR9 (right panel). Cells were pulsed with <t>BrdU</t> for 20 min and cell cycle analysis was performed after BrdU and <t>7-AAD</t> staining by flow cytometry. ( c ) HNSCC 136 PLVUT'-GFP (left panel) and HNSCC 136 PLVUT'-TLR9 (right panel) were induced with doxycycline and serum deprivated for 2 days. Cells lysate were collected 0, 4, 8 or 24 h after serum addition. Expression of cell cycle proteins were analyzed by western blotting. ( d ) Densitometry analysis of the western blot band from ( c ).
    Anti Brdu Ab, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Becton Dickinson anti brdu antibody
    Methylation kinetics of newly synthesized <t>DNA</t> at CI-f and CII-d. ES cells containing the genes for the indicated DNA methyltransferases were pulsed for 1 h with <t>BrdU.</t> The pulse was then removed, and fresh medium was added to the cells during the chase period. DNA was extracted from the cells at various times after the pulse began and was immunoprecipitated to isolate the BrdU-containing DNA. The methylation statuses of Hpa II sites in CI-f and CII-d at the indicated time points were determined by quantitative Ms-SNuPE analysis. Data are mean values from two to six experiments; error bars, standard deviations.
    Anti Brdu Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 2802 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti brdu antibody
    Dpp6 over-expression affects cell proliferation and apoptosis in RA induced P19 cells. A, Photograph showing <t>BrdU</t> incorporation in control (top) and Dpp6 over-expressing (bottom) P19 cells after 2 days of RA treatment and further culture for 2 days without RA. Cells were incubated with BrdU for 4 hrs, fixed and <t>immunostained</t> using antibody for BrdU while nuclei were detected using DAPI. B, Graph showing % of cells labeled with BrdU in control and Dpp6 expressing P19 cells after 4 days of differentiation. At least 500 cells were counted from each group. C D, Measurement of apoptosis in control and Dpp6 expressing P19 cells after 2 days of RA treatment and further culture for 4 days without RA by labeling with Annexin V-FITC and propidium iodide. % apoptotic cells were quantified using flow cytometry analysis. Values represent mean of three independent experiments and error bars represent ± SEM. *P
    Anti Brdu Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 1145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    BioLegend biotin anti brdu ab
    Dpp6 over-expression affects cell proliferation and apoptosis in RA induced P19 cells. A, Photograph showing <t>BrdU</t> incorporation in control (top) and Dpp6 over-expressing (bottom) P19 cells after 2 days of RA treatment and further culture for 2 days without RA. Cells were incubated with BrdU for 4 hrs, fixed and <t>immunostained</t> using antibody for BrdU while nuclei were detected using DAPI. B, Graph showing % of cells labeled with BrdU in control and Dpp6 expressing P19 cells after 4 days of differentiation. At least 500 cells were counted from each group. C D, Measurement of apoptosis in control and Dpp6 expressing P19 cells after 2 days of RA treatment and further culture for 4 days without RA by labeling with Annexin V-FITC and propidium iodide. % apoptotic cells were quantified using flow cytometry analysis. Values represent mean of three independent experiments and error bars represent ± SEM. *P
    Biotin Anti Brdu Ab, supplied by BioLegend, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Bio-Rad anti brdu primary ab
    IL-11 + Cells Appear in the Colon of DSS-Treated Mice and Express Stromal Cell Marker. a Il11-Egfp reporter mice were treated with 1.5% DSS in drinking water for 5 days, followed by a change to regular water. On day 7 after DSS treatment, Il11 and Egfp mRNA expression in the colon was determined by qPCR. Results are mean ± SE (n = 9 mice). b, c Appearance of IL-11 + cells in submucosal tissues of the colon of Il11-Egfp reporter mice on post-DSS treatment day 5 (b) or day 1 ( c ). Colonic tissue sections from untreated or DSS-treated Il11-Egfp reporter mice were H E stained or immunostained with <t>anti-GFP</t> antibody. Right panels show enlargements of the boxes (b). Scale bar, 100 μm. d, e Characterization of cell surface markers on IL-11 + cells. Colonic cells were prepared from the colon of Il11-Egfp reporter mice as in ( b ). We determined the percentages of EGFP + (IL-11 + ) cells from the colon before and after DSS treatment ( d ). Cells were stained with the indicated antibodies, and marker expressions were analyzed on GFP-positive cells ( e ). Results are representative of three independent experiments. f Representative immunostaining of IL-11 + cells. Colonic tissue sections were prepared from Il11-Egfp reporter mice as in ( b ), and immunostained with the indicated antibodies and anti-GFP antibody. Results are merged images. Right panels are enlarged images from the boxes (n = 3–4 mice). White arrowheads indicate merged cells. g IL-11 + cells do not proliferate in situ . Il11-Egfp reporter mice were treated with DSS as in ( a ), and intraperitoneally administered <t>BrdU</t> (40 mg/kg) on day 6. On day 7, colonic sections were prepared and stained with anti-GFP and anti-BrdU antibodies. Results are representative images from three independent experiments. Scale bars, 100 μm, unless otherwise indicated. Statistical significance was determined by two-tailed unpaired Student’s t -test ( a ). *p
    Anti Brdu Primary Ab, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore anti brdu
    <t>MARVELD1</t> affected granule cell migration but not proliferation. a Sagittal paraffin-embedded tissue sections of 0- and 6-day-old mice stained with HE. The whole cerebellum with low magnification showed the overall situation of abnormal cells. The arrowheads indicated migrating neurons. b In 6-day-old mice, the width of the EGL and the number of migrating granule cells were analyzed. c Granule cell proliferation and migration were evaluated after a short 1.5-h and a long 30-h chase following <t>BrdU</t> administration in 6-day-old mice in control and MARVELD1 KO animals. The relative cell number was counted. a – c : n = 3 for each genotype. ** p
    Anti Brdu, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Agilent technologies anti brdu ab
    Birth date analyses of <t>GFP-positive</t> cells in the cerebral cortical MZ by the <t>BrdU</t> labeling study. a, Quantitative analyses of the proportion of the number of BrdU and GFP coexpressing cells in the total number of GFP-expressing cells in E18.5 cerebral cortical MZ. Data points represent mean values ± SE from > 160 cells. Neocortical MZ was divided into three areas along the dorsolateral axis. There was no significant regional difference in the generation of GFP-positive cells among the three regions. The temporal pattern of generation of GFP-positive cells at the cingulate cortex was similar to that of neocortex. All the BrdU-labeled cells, including both the heavily labeled and weakly labeled cells, were counted for the analyses. b, ) for the detailed results of quantitative analyses. The percentage of GFP-positive cells generated in the three distinct regions of neocortex from E10.5 onward was 93.3% on average; the percentage of GFP-positive cells generated from E11.5 onward was 40.3% on average; the percentage of GFP-positive cells generated from E12.5 onward was 11.3% on average, the percentage of GFP-positive cells generated from E13.5 onward was 3.2% on average. By subtracting these numbers, we obtain, for example, 53% for the percentage of GFP-positive cells that are generated between E10.5 and E11.5 intervals. c, A representative coronal section showing the staining patterns of GFP and BrdU. A few number of GFP-, BrdU-double positive cells are shown. Scale bar, 20 μm.
    Anti Brdu Ab, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BioLegend anti brdu apc
    Birth date analyses of <t>GFP-positive</t> cells in the cerebral cortical MZ by the <t>BrdU</t> labeling study. a, Quantitative analyses of the proportion of the number of BrdU and GFP coexpressing cells in the total number of GFP-expressing cells in E18.5 cerebral cortical MZ. Data points represent mean values ± SE from > 160 cells. Neocortical MZ was divided into three areas along the dorsolateral axis. There was no significant regional difference in the generation of GFP-positive cells among the three regions. The temporal pattern of generation of GFP-positive cells at the cingulate cortex was similar to that of neocortex. All the BrdU-labeled cells, including both the heavily labeled and weakly labeled cells, were counted for the analyses. b, ) for the detailed results of quantitative analyses. The percentage of GFP-positive cells generated in the three distinct regions of neocortex from E10.5 onward was 93.3% on average; the percentage of GFP-positive cells generated from E11.5 onward was 40.3% on average; the percentage of GFP-positive cells generated from E12.5 onward was 11.3% on average, the percentage of GFP-positive cells generated from E13.5 onward was 3.2% on average. By subtracting these numbers, we obtain, for example, 53% for the percentage of GFP-positive cells that are generated between E10.5 and E11.5 intervals. c, A representative coronal section showing the staining patterns of GFP and BrdU. A few number of GFP-, BrdU-double positive cells are shown. Scale bar, 20 μm.
    Anti Brdu Apc, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti brdu mabs
    Profilerative index of wild-type and α1-null embryonic fibroblasts in vitro. ( A ) EFs were plated in presence of 2% FCS onto dishes uncoated or coated with fibrinogen (10 μg/ml), collagen I (100 μg/ml), or a mixture of collagen I (100 μg/ml) and collagen IV (30 μg/ml). 24 h after plating, cells were labeled with 10 μM <t>BrdU</t> ( Sigma Chemical Co. ), and incubated for a further 24 h. After staining with anti-BrdU <t>mAbs,</t> the Brdu labeling index (positive cells/total number of counted cells × 100) was determined by random evaluation of five different 40× microscopic fields for each duplicate sample, counting a minimum of 200 cells. Bars and errors indicate the mean and standard deviation. The differences seen between wild-type and α1-null groups on collagens are significant with P
    Anti Brdu Mabs, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    GH treatment promotes the proliferation of SGZ neurospheres. A) Neurospheres growing in defined media were treated for 24 h with GH (500 ng/mL), pegvisomant (Peg, 20 μg/mL), or GH + pegvisomant. Control cells were treated with saline. Four hours before the end of the treatment period cells were given a BrdU pulse (10 μM). Neurospheres were then dissociated, cells were collected by centrifugation onto coated cover slips, and BrdU was detected by immunocytochemistry. Each bar represents the mean + SEM of 3 experiments in triplicate. * = p

    Journal: BMC Neuroscience

    Article Title: Growth hormone pathways signaling for cell proliferation and survival in hippocampal neural precursors from postnatal mice

    doi: 10.1186/1471-2202-15-100

    Figure Lengend Snippet: GH treatment promotes the proliferation of SGZ neurospheres. A) Neurospheres growing in defined media were treated for 24 h with GH (500 ng/mL), pegvisomant (Peg, 20 μg/mL), or GH + pegvisomant. Control cells were treated with saline. Four hours before the end of the treatment period cells were given a BrdU pulse (10 μM). Neurospheres were then dissociated, cells were collected by centrifugation onto coated cover slips, and BrdU was detected by immunocytochemistry. Each bar represents the mean + SEM of 3 experiments in triplicate. * = p

    Article Snippet: SGZ cultures were then incubated overnight at 4°C with a primary anti-BrdU antibody (EMD Millipore Corporation; dilution 1:50) in PBS containing 0.1% Triton X-100 and 0.3% BSA.

    Techniques: Centrifugation, Immunocytochemistry

    The receptor RAGE critically mediates the cellular response to S100A8/A9. a: Direct blockade by a specific RAGE blocking antibody reduces cellular proliferation. RAGE dependent proliferation was analyzed in normal primary, AK and SCC cells. Cells were incubated for 1 hour with a blocking anti-RAGE antibody (80μg/ml, as recommended by manufacturer) followed by S100A8/A9 stimulation for additional 24 hours. The differences in the proliferation after the blockade were assessed by BrdU incorporation (1 way Anova, Bonferroni`s Multiple test, **p

    Journal: PLoS ONE

    Article Title: S100A8/A9 Stimulates Keratinocyte Proliferation in the Development of Squamous Cell Carcinoma of the Skin via the Receptor for Advanced Glycation-End Products

    doi: 10.1371/journal.pone.0120971

    Figure Lengend Snippet: The receptor RAGE critically mediates the cellular response to S100A8/A9. a: Direct blockade by a specific RAGE blocking antibody reduces cellular proliferation. RAGE dependent proliferation was analyzed in normal primary, AK and SCC cells. Cells were incubated for 1 hour with a blocking anti-RAGE antibody (80μg/ml, as recommended by manufacturer) followed by S100A8/A9 stimulation for additional 24 hours. The differences in the proliferation after the blockade were assessed by BrdU incorporation (1 way Anova, Bonferroni`s Multiple test, **p

    Article Snippet: Double staining for RAGE and BrdU was performed using the specific RAGE antibody and specific mouse anti BrdU antibody (Millipore ).

    Techniques: Blocking Assay, Incubation, BrdU Incorporation Assay

    Endogenous S100A8/A9 is involved in cellular proliferation. a: Spontaneous secretion of S100A8/A9 from normal and SCC-derived keratinocytes. Normal and SCC-derived keratinocytes were grown in 96 well plates for 24 hours. Afterwards, supernatant was collected and preceded for the assessment of secreted S100A8/A9 using specific ELISA for S100A8/A9. b: Blockade of RAGE using anti RAGE blocking antibody reduces spontaneous proliferation of keratinocytes. Normal and SCC-derived keratinocytes were incubated with a blocking anti-RAGE antibody (8ug/100μl) for 24 hours. A decrease of the proliferation rate was detected (20–30%) based on the BrdU incorporation (t-test *p = 0.002). All the results are presented as percentage deviation from corresponding control and represent the mean +/- SD of duplicate values.

    Journal: PLoS ONE

    Article Title: S100A8/A9 Stimulates Keratinocyte Proliferation in the Development of Squamous Cell Carcinoma of the Skin via the Receptor for Advanced Glycation-End Products

    doi: 10.1371/journal.pone.0120971

    Figure Lengend Snippet: Endogenous S100A8/A9 is involved in cellular proliferation. a: Spontaneous secretion of S100A8/A9 from normal and SCC-derived keratinocytes. Normal and SCC-derived keratinocytes were grown in 96 well plates for 24 hours. Afterwards, supernatant was collected and preceded for the assessment of secreted S100A8/A9 using specific ELISA for S100A8/A9. b: Blockade of RAGE using anti RAGE blocking antibody reduces spontaneous proliferation of keratinocytes. Normal and SCC-derived keratinocytes were incubated with a blocking anti-RAGE antibody (8ug/100μl) for 24 hours. A decrease of the proliferation rate was detected (20–30%) based on the BrdU incorporation (t-test *p = 0.002). All the results are presented as percentage deviation from corresponding control and represent the mean +/- SD of duplicate values.

    Article Snippet: Double staining for RAGE and BrdU was performed using the specific RAGE antibody and specific mouse anti BrdU antibody (Millipore ).

    Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Incubation, BrdU Incorporation Assay, T-Test

    Intranasal HB-EGF accelerates oligodendrocyte regeneration and promotes cellular recovery in white matter after neonatal hypoxia a, Confocal image of a preterm corpus callosum (CC) highlighting an Olig2 + EGFR + cell (box). b, Protocol of intranasal HB-EGF/BrdU administration and tissue collection. c, Number of WM Rep + Olig2 + and Rep + CC1 + cells in HB-EGF-treated mice. d, HB-EGF attenuated or prevented the effects of Hyp on OL apoptosis. e, HB-EGF had an additive effect on Hyp-induced increase of Rep + NG2 + OPCs at P15, but not at P18. f, HB-EGF had an additive effect on Hyp-induced increase of OPC proliferation. g, HB-EGF promoted oligodendrogenesis after Hyp at P18. h, Fate-mapping of OPCs [PDGFαR-CreER;Z/EG (GFP) mouse] demonstrated that oligodendrogenesis occurred from PDGFαR + cells. c–h, n=4 mice per group and per age; One-way ANOVA, Bonferroni post hoc test for individual comparisons. i. Removal of EGFR in PDGFαR-expressing OPCs (PDGFαR-CreER;EGFR fl/fl ;Z/EG mouse) caused a decrease in NG2 + OPCs after Hyp and prevented the effects of HB-EGF (n=4 mice per group except Hyp HB-EGF PDGFαR-CreER;EGFR fl/fl n=3; One-way ANOVA of all 4 groups with a post hoc unpaired t-tests). j, HB-EGF promoted recovery in WM MBP and PLP protein levels after Hyp (Western blot) (n=6 mice per group except Nx HB-EGF n=4; One-way ANOVA, Bonferroni post hoc test for individual comparisons). All histograms are presented as means ± s.e.m. ^P=0.05; *P

    Journal: Nature

    Article Title: Intranasal epidermal growth factor treatment rescues neonatal brain injury

    doi: 10.1038/nature12880

    Figure Lengend Snippet: Intranasal HB-EGF accelerates oligodendrocyte regeneration and promotes cellular recovery in white matter after neonatal hypoxia a, Confocal image of a preterm corpus callosum (CC) highlighting an Olig2 + EGFR + cell (box). b, Protocol of intranasal HB-EGF/BrdU administration and tissue collection. c, Number of WM Rep + Olig2 + and Rep + CC1 + cells in HB-EGF-treated mice. d, HB-EGF attenuated or prevented the effects of Hyp on OL apoptosis. e, HB-EGF had an additive effect on Hyp-induced increase of Rep + NG2 + OPCs at P15, but not at P18. f, HB-EGF had an additive effect on Hyp-induced increase of OPC proliferation. g, HB-EGF promoted oligodendrogenesis after Hyp at P18. h, Fate-mapping of OPCs [PDGFαR-CreER;Z/EG (GFP) mouse] demonstrated that oligodendrogenesis occurred from PDGFαR + cells. c–h, n=4 mice per group and per age; One-way ANOVA, Bonferroni post hoc test for individual comparisons. i. Removal of EGFR in PDGFαR-expressing OPCs (PDGFαR-CreER;EGFR fl/fl ;Z/EG mouse) caused a decrease in NG2 + OPCs after Hyp and prevented the effects of HB-EGF (n=4 mice per group except Hyp HB-EGF PDGFαR-CreER;EGFR fl/fl n=3; One-way ANOVA of all 4 groups with a post hoc unpaired t-tests). j, HB-EGF promoted recovery in WM MBP and PLP protein levels after Hyp (Western blot) (n=6 mice per group except Nx HB-EGF n=4; One-way ANOVA, Bonferroni post hoc test for individual comparisons). All histograms are presented as means ± s.e.m. ^P=0.05; *P

    Article Snippet: Primary antibody dilutions were 1:500 for anti-BrdU (Accurate), anti-NG2 (Millipore), anti-Olig2 (Millipore), anti-Ki67 (Vector), anti-APC (also referred as CC1, Millipore) and anti-cleaved caspase-3 (caspase-3; Millipore); 1:250 for anti-MBP (Covance); and 1:500 anti-EGFR phosphorylated Tyr1068 (Novus Biologicals).

    Techniques: Mouse Assay, Expressing, Plasmid Purification, Western Blot

    EGFR activity is crucial for white matter recovery after neonatal hypoxia a, Protocol of Gefitinib and BrdU administration. b, Western blot of WM shows that Gefitinib decreased pEGFR in Nx and prevented the increase in pEGFR after Hyp (n=5 mice per group; One-way ANOVA, Bonferroni post hoc test for individual comparisons). c, Counts of Rep + Olig2 + and Rep + CC1 + cells in WM. d, Gerfinitib increased cell apoptosis in Nx and Hyp. e,f, Gefitinib decreased Rep + NG2 + OPCs (e) and OL-lineage cell proliferation (f) in Nx and prevented Hyp-induced increase in OPC and OL-lineage cell proliferation. g, Lon-term effects of Gefinitib on Rep + Olig2 + and Rep + CC1 + cells. h, Gefinitib decreased newly-generated Rep + CC1 + OLs in Nx and prevented oligodendrogenesis after Hyp. i, Gefitinib prevented recovery of CNPase and MBP expression after Hyp. c–i; n=4 mice per all groups and per age; One-way ANOVA, Bonferroni post hoc test for individual comparisons. All histograms are presented as means ± s.e.m. ^P=0.05; *P

    Journal: Nature

    Article Title: Intranasal epidermal growth factor treatment rescues neonatal brain injury

    doi: 10.1038/nature12880

    Figure Lengend Snippet: EGFR activity is crucial for white matter recovery after neonatal hypoxia a, Protocol of Gefitinib and BrdU administration. b, Western blot of WM shows that Gefitinib decreased pEGFR in Nx and prevented the increase in pEGFR after Hyp (n=5 mice per group; One-way ANOVA, Bonferroni post hoc test for individual comparisons). c, Counts of Rep + Olig2 + and Rep + CC1 + cells in WM. d, Gerfinitib increased cell apoptosis in Nx and Hyp. e,f, Gefitinib decreased Rep + NG2 + OPCs (e) and OL-lineage cell proliferation (f) in Nx and prevented Hyp-induced increase in OPC and OL-lineage cell proliferation. g, Lon-term effects of Gefinitib on Rep + Olig2 + and Rep + CC1 + cells. h, Gefinitib decreased newly-generated Rep + CC1 + OLs in Nx and prevented oligodendrogenesis after Hyp. i, Gefitinib prevented recovery of CNPase and MBP expression after Hyp. c–i; n=4 mice per all groups and per age; One-way ANOVA, Bonferroni post hoc test for individual comparisons. All histograms are presented as means ± s.e.m. ^P=0.05; *P

    Article Snippet: Primary antibody dilutions were 1:500 for anti-BrdU (Accurate), anti-NG2 (Millipore), anti-Olig2 (Millipore), anti-Ki67 (Vector), anti-APC (also referred as CC1, Millipore) and anti-cleaved caspase-3 (caspase-3; Millipore); 1:250 for anti-MBP (Covance); and 1:500 anti-EGFR phosphorylated Tyr1068 (Novus Biologicals).

    Techniques: Activity Assay, Western Blot, Mouse Assay, Generated, Expressing

    TLR9 expression affected the cell cycle but not apoptosis. ( a ) Caski cells stably transduced with GFP (left panel) or TLR9 (right panel) encoded pbabe were stained with Annexin V and propidium iodide to determine percentage of apoptotic and necrotic cells. ( b ) HNSCC 136 cells were stably transduced with pbabe (left panel), pbabe -GFP (middle panel) or pbabe-TLR9 (right panel). Cells were pulsed with BrdU for 20 min and cell cycle analysis was performed after BrdU and 7-AAD staining by flow cytometry. ( c ) HNSCC 136 PLVUT'-GFP (left panel) and HNSCC 136 PLVUT'-TLR9 (right panel) were induced with doxycycline and serum deprivated for 2 days. Cells lysate were collected 0, 4, 8 or 24 h after serum addition. Expression of cell cycle proteins were analyzed by western blotting. ( d ) Densitometry analysis of the western blot band from ( c ).

    Journal: Oncogenesis

    Article Title: TLR9 re-expression in cancer cells extends the S-phase and stabilizes p16INK4a protein expression

    doi: 10.1038/oncsis.2016.49

    Figure Lengend Snippet: TLR9 expression affected the cell cycle but not apoptosis. ( a ) Caski cells stably transduced with GFP (left panel) or TLR9 (right panel) encoded pbabe were stained with Annexin V and propidium iodide to determine percentage of apoptotic and necrotic cells. ( b ) HNSCC 136 cells were stably transduced with pbabe (left panel), pbabe -GFP (middle panel) or pbabe-TLR9 (right panel). Cells were pulsed with BrdU for 20 min and cell cycle analysis was performed after BrdU and 7-AAD staining by flow cytometry. ( c ) HNSCC 136 PLVUT'-GFP (left panel) and HNSCC 136 PLVUT'-TLR9 (right panel) were induced with doxycycline and serum deprivated for 2 days. Cells lysate were collected 0, 4, 8 or 24 h after serum addition. Expression of cell cycle proteins were analyzed by western blotting. ( d ) Densitometry analysis of the western blot band from ( c ).

    Article Snippet: The cells were then blocked and stained with an anti-BrdU (Biolegend, London, UK) and 7-AAD (Life Technologies).

    Techniques: Expressing, Stable Transfection, Transduction, Staining, Cell Cycle Assay, Flow Cytometry, Cytometry, Western Blot

    Methylation kinetics of newly synthesized DNA at CI-f and CII-d. ES cells containing the genes for the indicated DNA methyltransferases were pulsed for 1 h with BrdU. The pulse was then removed, and fresh medium was added to the cells during the chase period. DNA was extracted from the cells at various times after the pulse began and was immunoprecipitated to isolate the BrdU-containing DNA. The methylation statuses of Hpa II sites in CI-f and CII-d at the indicated time points were determined by quantitative Ms-SNuPE analysis. Data are mean values from two to six experiments; error bars, standard deviations.

    Journal: Molecular and Cellular Biology

    Article Title: Cooperativity between DNA Methyltransferases in the Maintenance Methylation of Repetitive Elements

    doi: 10.1128/MCB.22.2.480-491.2002

    Figure Lengend Snippet: Methylation kinetics of newly synthesized DNA at CI-f and CII-d. ES cells containing the genes for the indicated DNA methyltransferases were pulsed for 1 h with BrdU. The pulse was then removed, and fresh medium was added to the cells during the chase period. DNA was extracted from the cells at various times after the pulse began and was immunoprecipitated to isolate the BrdU-containing DNA. The methylation statuses of Hpa II sites in CI-f and CII-d at the indicated time points were determined by quantitative Ms-SNuPE analysis. Data are mean values from two to six experiments; error bars, standard deviations.

    Article Snippet: The samples were denatured at 95°C for 5 min, cooled on ice for 2 min, and immediately mixed with 2.4 μl of anti-BrdU antibody (25 μg/ml) per μg of DNA (Becton Dickinson) ( ).

    Techniques: Methylation, Synthesized, Immunoprecipitation, Mass Spectrometry

    Dpp6 over-expression affects cell proliferation and apoptosis in RA induced P19 cells. A, Photograph showing BrdU incorporation in control (top) and Dpp6 over-expressing (bottom) P19 cells after 2 days of RA treatment and further culture for 2 days without RA. Cells were incubated with BrdU for 4 hrs, fixed and immunostained using antibody for BrdU while nuclei were detected using DAPI. B, Graph showing % of cells labeled with BrdU in control and Dpp6 expressing P19 cells after 4 days of differentiation. At least 500 cells were counted from each group. C D, Measurement of apoptosis in control and Dpp6 expressing P19 cells after 2 days of RA treatment and further culture for 4 days without RA by labeling with Annexin V-FITC and propidium iodide. % apoptotic cells were quantified using flow cytometry analysis. Values represent mean of three independent experiments and error bars represent ± SEM. *P

    Journal: PLoS ONE

    Article Title: Epigenetic Regulation of Dpp6 Expression by Dnmt3b and Its Novel Role in the Inhibition of RA Induced Neuronal Differentiation of P19 Cells

    doi: 10.1371/journal.pone.0055826

    Figure Lengend Snippet: Dpp6 over-expression affects cell proliferation and apoptosis in RA induced P19 cells. A, Photograph showing BrdU incorporation in control (top) and Dpp6 over-expressing (bottom) P19 cells after 2 days of RA treatment and further culture for 2 days without RA. Cells were incubated with BrdU for 4 hrs, fixed and immunostained using antibody for BrdU while nuclei were detected using DAPI. B, Graph showing % of cells labeled with BrdU in control and Dpp6 expressing P19 cells after 4 days of differentiation. At least 500 cells were counted from each group. C D, Measurement of apoptosis in control and Dpp6 expressing P19 cells after 2 days of RA treatment and further culture for 4 days without RA by labeling with Annexin V-FITC and propidium iodide. % apoptotic cells were quantified using flow cytometry analysis. Values represent mean of three independent experiments and error bars represent ± SEM. *P

    Article Snippet: Cells were then fixed, denatured and immunostained with anti-BrdU antibody (Abcam).

    Techniques: Over Expression, BrdU Incorporation Assay, Expressing, Incubation, Labeling, Flow Cytometry, Cytometry

    Metformin rejuvenation of neurogenic potential in aged mice. (A) The subventricular zone (SVZ) of old mice treated with metformin ( n ≥ 3) immunostained with the neural stem cell marker, Sox2, and newborn neuronal marker, GFAP. Right: Quantification of (B) Sox2 + cells and (C) DCX + cells. (D) SVZ of old mice immunostained with the neuronal cell marker, NeuN, and cell proliferation marker, BrdU. Right: Quantification of (E) BrdU + /NeuN + cells to quantify newborn neurons. (F) Representative fields of GFAP and Tuj1 immunofluorescence staining of cultured neural stem cells treated with metformin after 7 days of spontaneous differentiation. Right: Statistical analysis of percentages of (G) GFAP + cells and (H) Tuj1 + cells. Scale bar = 100 μm. Ctrl: Control; Met: Metformin. The overall significance between two groups was determined by Student’s t -test. * p

    Journal: bioRxiv

    Article Title: Metformin improves cognition of aged mice by promoting cerebral angiogenesis and neurogenesis

    doi: 10.1101/2020.03.25.006767

    Figure Lengend Snippet: Metformin rejuvenation of neurogenic potential in aged mice. (A) The subventricular zone (SVZ) of old mice treated with metformin ( n ≥ 3) immunostained with the neural stem cell marker, Sox2, and newborn neuronal marker, GFAP. Right: Quantification of (B) Sox2 + cells and (C) DCX + cells. (D) SVZ of old mice immunostained with the neuronal cell marker, NeuN, and cell proliferation marker, BrdU. Right: Quantification of (E) BrdU + /NeuN + cells to quantify newborn neurons. (F) Representative fields of GFAP and Tuj1 immunofluorescence staining of cultured neural stem cells treated with metformin after 7 days of spontaneous differentiation. Right: Statistical analysis of percentages of (G) GFAP + cells and (H) Tuj1 + cells. Scale bar = 100 μm. Ctrl: Control; Met: Metformin. The overall significance between two groups was determined by Student’s t -test. * p

    Article Snippet: Primary antibodies used in this study include: anti-GFAP (1:1000; Dako, Glostrup, Denmark), anti-Tuj1 (1:500; R & D Systems, Minneapolis, MN, USA), anti-DCX (1:400; Cell Signaling Technology, Danvers, MA, USA), anti-BrdU (1:200; Abcam, Cambridge, UK), anti-Sox2 (1:200, Abcam), and anti-NeuN (1:300; Abcam).

    Techniques: Mouse Assay, Marker, Immunofluorescence, Staining, Cell Culture

    IL-11 + Cells Appear in the Colon of DSS-Treated Mice and Express Stromal Cell Marker. a Il11-Egfp reporter mice were treated with 1.5% DSS in drinking water for 5 days, followed by a change to regular water. On day 7 after DSS treatment, Il11 and Egfp mRNA expression in the colon was determined by qPCR. Results are mean ± SE (n = 9 mice). b, c Appearance of IL-11 + cells in submucosal tissues of the colon of Il11-Egfp reporter mice on post-DSS treatment day 5 (b) or day 1 ( c ). Colonic tissue sections from untreated or DSS-treated Il11-Egfp reporter mice were H E stained or immunostained with anti-GFP antibody. Right panels show enlargements of the boxes (b). Scale bar, 100 μm. d, e Characterization of cell surface markers on IL-11 + cells. Colonic cells were prepared from the colon of Il11-Egfp reporter mice as in ( b ). We determined the percentages of EGFP + (IL-11 + ) cells from the colon before and after DSS treatment ( d ). Cells were stained with the indicated antibodies, and marker expressions were analyzed on GFP-positive cells ( e ). Results are representative of three independent experiments. f Representative immunostaining of IL-11 + cells. Colonic tissue sections were prepared from Il11-Egfp reporter mice as in ( b ), and immunostained with the indicated antibodies and anti-GFP antibody. Results are merged images. Right panels are enlarged images from the boxes (n = 3–4 mice). White arrowheads indicate merged cells. g IL-11 + cells do not proliferate in situ . Il11-Egfp reporter mice were treated with DSS as in ( a ), and intraperitoneally administered BrdU (40 mg/kg) on day 6. On day 7, colonic sections were prepared and stained with anti-GFP and anti-BrdU antibodies. Results are representative images from three independent experiments. Scale bars, 100 μm, unless otherwise indicated. Statistical significance was determined by two-tailed unpaired Student’s t -test ( a ). *p

    Journal: bioRxiv

    Article Title: Interleukin-11 is a Marker for Both Cancer- and Inflammation-Associated Fibroblasts that Contribute to Colorectal Cancer Progression

    doi: 10.1101/2020.01.25.919795

    Figure Lengend Snippet: IL-11 + Cells Appear in the Colon of DSS-Treated Mice and Express Stromal Cell Marker. a Il11-Egfp reporter mice were treated with 1.5% DSS in drinking water for 5 days, followed by a change to regular water. On day 7 after DSS treatment, Il11 and Egfp mRNA expression in the colon was determined by qPCR. Results are mean ± SE (n = 9 mice). b, c Appearance of IL-11 + cells in submucosal tissues of the colon of Il11-Egfp reporter mice on post-DSS treatment day 5 (b) or day 1 ( c ). Colonic tissue sections from untreated or DSS-treated Il11-Egfp reporter mice were H E stained or immunostained with anti-GFP antibody. Right panels show enlargements of the boxes (b). Scale bar, 100 μm. d, e Characterization of cell surface markers on IL-11 + cells. Colonic cells were prepared from the colon of Il11-Egfp reporter mice as in ( b ). We determined the percentages of EGFP + (IL-11 + ) cells from the colon before and after DSS treatment ( d ). Cells were stained with the indicated antibodies, and marker expressions were analyzed on GFP-positive cells ( e ). Results are representative of three independent experiments. f Representative immunostaining of IL-11 + cells. Colonic tissue sections were prepared from Il11-Egfp reporter mice as in ( b ), and immunostained with the indicated antibodies and anti-GFP antibody. Results are merged images. Right panels are enlarged images from the boxes (n = 3–4 mice). White arrowheads indicate merged cells. g IL-11 + cells do not proliferate in situ . Il11-Egfp reporter mice were treated with DSS as in ( a ), and intraperitoneally administered BrdU (40 mg/kg) on day 6. On day 7, colonic sections were prepared and stained with anti-GFP and anti-BrdU antibodies. Results are representative images from three independent experiments. Scale bars, 100 μm, unless otherwise indicated. Statistical significance was determined by two-tailed unpaired Student’s t -test ( a ). *p

    Article Snippet: The following antibodies used in this study were obtained from the indicated sources: anti-phospho-ERK (4370, CST), anti-Ki67 (ab16667, Abcam), anti-GFP (GFP-Go-Af1480 or GFP-Rb-Af2020, Frontier Institute), anti-BrdU (BU1/75, BIO-RAD), anti-IL-11 (LS-C408373, LSBio), anti-CD45 (13917, CST), anti-CD45 (IR751, Dako), anti-podoplanin (127403, BioLegend), anti-α-SMA (ab5694, Abcam), anti-collagen I (ab34710, Abcam), anti-collagen IV (ab6586, Abcam), anti-E-cadherin (560062, BD Biosciences), anti-E-cadherin (NCH-38, Dako), anti-vimentin (9856, CST), anti-phospho-STAT3 (9145, CST), anti-STAT3 (SC-482, Santa Cruz), anti-β-Actin (622102, Biolegend), and anti-tubulin (T5168, Sigma-Aldrich).

    Techniques: Mouse Assay, Marker, Expressing, Real-time Polymerase Chain Reaction, Staining, Immunostaining, In Situ, Two Tailed Test

    MARVELD1 affected granule cell migration but not proliferation. a Sagittal paraffin-embedded tissue sections of 0- and 6-day-old mice stained with HE. The whole cerebellum with low magnification showed the overall situation of abnormal cells. The arrowheads indicated migrating neurons. b In 6-day-old mice, the width of the EGL and the number of migrating granule cells were analyzed. c Granule cell proliferation and migration were evaluated after a short 1.5-h and a long 30-h chase following BrdU administration in 6-day-old mice in control and MARVELD1 KO animals. The relative cell number was counted. a – c : n = 3 for each genotype. ** p

    Journal: Cell Death & Disease

    Article Title: MARVELD1 depletion leads to dysfunction of motor and cognition via regulating glia-dependent neuronal migration during brain development

    doi: 10.1038/s41419-018-1027-6

    Figure Lengend Snippet: MARVELD1 affected granule cell migration but not proliferation. a Sagittal paraffin-embedded tissue sections of 0- and 6-day-old mice stained with HE. The whole cerebellum with low magnification showed the overall situation of abnormal cells. The arrowheads indicated migrating neurons. b In 6-day-old mice, the width of the EGL and the number of migrating granule cells were analyzed. c Granule cell proliferation and migration were evaluated after a short 1.5-h and a long 30-h chase following BrdU administration in 6-day-old mice in control and MARVELD1 KO animals. The relative cell number was counted. a – c : n = 3 for each genotype. ** p

    Article Snippet: For immunohistochemistry and immunofluorescence, the following primary antibodies were used: anti-Calb (1:200, Sigma, no.sab4200543), anti-Blbp (1:100, Abcam, no.ab32423), anti-GFAP (1:200, Abcam, no.ab7260 or ab10062), anti-ITGB1 (1:100, Abcam, no.ab183666 or ab95623), anti-FAK (1:100, Abcam, no.ab40794), anti-p397-FAK (1:100, Abcam, no.ab81298), anti-BrdU (1:100, Sigma, no.b2531), anti-MARVELD1 (1:100, Abcam, no.ab91640 or no.ab169184) and anti-NeuN (1:200, Abcam, no.ab104224).

    Techniques: Migration, Mouse Assay, Staining

    MARVELD1 regulated accurate radial migration by affecting the formation of glial fibres. a Immunofluorescence staining of MARVELD1 (green) and glial cells marker GFAP (red) in 6-day-old mice cerebellum. The arrowheads indicated glial cells. b HE staining of 4-week-old GFAP-cre/MARVELD1 fl/fl mice cerebellum sections. The whole cerebellum with low magnification showed the overall situation of abnormal cells location in the molecular layer. n = 3 for each genotype. a and b were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. c HE staining of 6-day-old control mice and GFAP-cre/MARVELD1 fl/fl mice cerebellum. The whole cerebellum with low magnification showed the overall situation of abnormal cells location in the molecular layer. n = 3 for each genotype. a and b were different areas from control cerebellum. c , d and e are different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. d Granule cell migration was evaluated by a long 60-h chase following BrdU administration in 6-day-old control mice and GFAP-cre/MARVELD1 fl/fl mice. n = 3 for each genotype. e Immunohistochemistry staining with GFAP antibodies in 4-week-old control and GFAP-cre/MARVELD1 fl/fl cerebella. n = 3 for each genotype. a and b were different areas from control cerebellum. c and d were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. f Immunohistochemistry staining with Calb antibodies in control and GFAP-cre/MARVELD1 fl/fl cerebella in 4-week-old mice. n = 3 for each genotype. a and b are different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. g Sagittal sections of 6-day-old mice cerebellum immunostained with anti-GFAP. GFAP-cre/MARVELD1 fl/fl mice had no obvious glial fibres. n = 3 for each genotype. a and b were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. h Immunofluorescence of Calb (red) in 6-day-old mice cerebellum. *** p

    Journal: Cell Death & Disease

    Article Title: MARVELD1 depletion leads to dysfunction of motor and cognition via regulating glia-dependent neuronal migration during brain development

    doi: 10.1038/s41419-018-1027-6

    Figure Lengend Snippet: MARVELD1 regulated accurate radial migration by affecting the formation of glial fibres. a Immunofluorescence staining of MARVELD1 (green) and glial cells marker GFAP (red) in 6-day-old mice cerebellum. The arrowheads indicated glial cells. b HE staining of 4-week-old GFAP-cre/MARVELD1 fl/fl mice cerebellum sections. The whole cerebellum with low magnification showed the overall situation of abnormal cells location in the molecular layer. n = 3 for each genotype. a and b were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. c HE staining of 6-day-old control mice and GFAP-cre/MARVELD1 fl/fl mice cerebellum. The whole cerebellum with low magnification showed the overall situation of abnormal cells location in the molecular layer. n = 3 for each genotype. a and b were different areas from control cerebellum. c , d and e are different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. d Granule cell migration was evaluated by a long 60-h chase following BrdU administration in 6-day-old control mice and GFAP-cre/MARVELD1 fl/fl mice. n = 3 for each genotype. e Immunohistochemistry staining with GFAP antibodies in 4-week-old control and GFAP-cre/MARVELD1 fl/fl cerebella. n = 3 for each genotype. a and b were different areas from control cerebellum. c and d were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. f Immunohistochemistry staining with Calb antibodies in control and GFAP-cre/MARVELD1 fl/fl cerebella in 4-week-old mice. n = 3 for each genotype. a and b are different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. g Sagittal sections of 6-day-old mice cerebellum immunostained with anti-GFAP. GFAP-cre/MARVELD1 fl/fl mice had no obvious glial fibres. n = 3 for each genotype. a and b were different areas from GFAP-cre/MARVELD1 fl/fl cerebellum. h Immunofluorescence of Calb (red) in 6-day-old mice cerebellum. *** p

    Article Snippet: For immunohistochemistry and immunofluorescence, the following primary antibodies were used: anti-Calb (1:200, Sigma, no.sab4200543), anti-Blbp (1:100, Abcam, no.ab32423), anti-GFAP (1:200, Abcam, no.ab7260 or ab10062), anti-ITGB1 (1:100, Abcam, no.ab183666 or ab95623), anti-FAK (1:100, Abcam, no.ab40794), anti-p397-FAK (1:100, Abcam, no.ab81298), anti-BrdU (1:100, Sigma, no.b2531), anti-MARVELD1 (1:100, Abcam, no.ab91640 or no.ab169184) and anti-NeuN (1:200, Abcam, no.ab104224).

    Techniques: Migration, Immunofluorescence, Staining, Marker, Mouse Assay, Immunohistochemistry

    Birth date analyses of GFP-positive cells in the cerebral cortical MZ by the BrdU labeling study. a, Quantitative analyses of the proportion of the number of BrdU and GFP coexpressing cells in the total number of GFP-expressing cells in E18.5 cerebral cortical MZ. Data points represent mean values ± SE from > 160 cells. Neocortical MZ was divided into three areas along the dorsolateral axis. There was no significant regional difference in the generation of GFP-positive cells among the three regions. The temporal pattern of generation of GFP-positive cells at the cingulate cortex was similar to that of neocortex. All the BrdU-labeled cells, including both the heavily labeled and weakly labeled cells, were counted for the analyses. b, ) for the detailed results of quantitative analyses. The percentage of GFP-positive cells generated in the three distinct regions of neocortex from E10.5 onward was 93.3% on average; the percentage of GFP-positive cells generated from E11.5 onward was 40.3% on average; the percentage of GFP-positive cells generated from E12.5 onward was 11.3% on average, the percentage of GFP-positive cells generated from E13.5 onward was 3.2% on average. By subtracting these numbers, we obtain, for example, 53% for the percentage of GFP-positive cells that are generated between E10.5 and E11.5 intervals. c, A representative coronal section showing the staining patterns of GFP and BrdU. A few number of GFP-, BrdU-double positive cells are shown. Scale bar, 20 μm.

    Journal: The Journal of Neuroscience

    Article Title: Generation of Reelin-Positive Marginal Zone Cells from the Caudomedial Wall of Telencephalic Vesicles

    doi: 10.1523/JNEUROSCI.4671-03.2004

    Figure Lengend Snippet: Birth date analyses of GFP-positive cells in the cerebral cortical MZ by the BrdU labeling study. a, Quantitative analyses of the proportion of the number of BrdU and GFP coexpressing cells in the total number of GFP-expressing cells in E18.5 cerebral cortical MZ. Data points represent mean values ± SE from > 160 cells. Neocortical MZ was divided into three areas along the dorsolateral axis. There was no significant regional difference in the generation of GFP-positive cells among the three regions. The temporal pattern of generation of GFP-positive cells at the cingulate cortex was similar to that of neocortex. All the BrdU-labeled cells, including both the heavily labeled and weakly labeled cells, were counted for the analyses. b, ) for the detailed results of quantitative analyses. The percentage of GFP-positive cells generated in the three distinct regions of neocortex from E10.5 onward was 93.3% on average; the percentage of GFP-positive cells generated from E11.5 onward was 40.3% on average; the percentage of GFP-positive cells generated from E12.5 onward was 11.3% on average, the percentage of GFP-positive cells generated from E13.5 onward was 3.2% on average. By subtracting these numbers, we obtain, for example, 53% for the percentage of GFP-positive cells that are generated between E10.5 and E11.5 intervals. c, A representative coronal section showing the staining patterns of GFP and BrdU. A few number of GFP-, BrdU-double positive cells are shown. Scale bar, 20 μm.

    Article Snippet: BrdU and GFP were detected using anti-BrdU Ab (Dako, Carpinteria, CA) and anti-GFP Ab as described ( ).

    Techniques: Labeling, Expressing, Generated, Staining

    Profilerative index of wild-type and α1-null embryonic fibroblasts in vitro. ( A ) EFs were plated in presence of 2% FCS onto dishes uncoated or coated with fibrinogen (10 μg/ml), collagen I (100 μg/ml), or a mixture of collagen I (100 μg/ml) and collagen IV (30 μg/ml). 24 h after plating, cells were labeled with 10 μM BrdU ( Sigma Chemical Co. ), and incubated for a further 24 h. After staining with anti-BrdU mAbs, the Brdu labeling index (positive cells/total number of counted cells × 100) was determined by random evaluation of five different 40× microscopic fields for each duplicate sample, counting a minimum of 200 cells. Bars and errors indicate the mean and standard deviation. The differences seen between wild-type and α1-null groups on collagens are significant with P

    Journal: The Journal of Cell Biology

    Article Title: Integrin ?1?1 Mediates a Unique Collagen-dependent Proliferation Pathway In Vivo

    doi:

    Figure Lengend Snippet: Profilerative index of wild-type and α1-null embryonic fibroblasts in vitro. ( A ) EFs were plated in presence of 2% FCS onto dishes uncoated or coated with fibrinogen (10 μg/ml), collagen I (100 μg/ml), or a mixture of collagen I (100 μg/ml) and collagen IV (30 μg/ml). 24 h after plating, cells were labeled with 10 μM BrdU ( Sigma Chemical Co. ), and incubated for a further 24 h. After staining with anti-BrdU mAbs, the Brdu labeling index (positive cells/total number of counted cells × 100) was determined by random evaluation of five different 40× microscopic fields for each duplicate sample, counting a minimum of 200 cells. Bars and errors indicate the mean and standard deviation. The differences seen between wild-type and α1-null groups on collagens are significant with P

    Article Snippet: After 24 h, cells were labeled with 10 μM 5′-bromo-2′-deoxy-uridine (BrdU; Sigma Chemical Co. ), and incubated for a further 24 h. Cells were then washed, fixed, and stained with anti-BrdU mAbs ( Sigma Chemical Co. ), followed by biotinylated secondary anti–mouse antibody and avidin/biotin-HRP–conjugated complex (Vector Laboratories, Burlingame, CA).

    Techniques: In Vitro, Labeling, Incubation, Staining, Standard Deviation