dna fragmentation  (Roche)


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    Structured Review

    Roche dna fragmentation
    Lung slides from WT and p66 Shc−/− mice scored at 5 months for active caspase-3, <t>TUNEL</t> and p16INK4a. Immunohistochemical evaluation of active caspase-3 on lungs of air control (A) and smoking WT (B) mice at 5 months. Note in (B) a positive staining on alveolar macrophages and epithelial lung cells. The positivity on lung epithelial cells is more evident at higher magnification (see the arrowheads in the inset). Active caspase-3 staining on tissue slides from air-control (C) and smoking p66 Shc−/− mice (D). <t>DNA</t> strand-break extremities labeling (TUNEL) on lung tissues from air-control (E) and smoking (F) WT at 5 months. In (G) and (H) TUNEL labeling on air-control and smoking p66 Shc−/− mice, respectively. Immunohistochemical staining for the cyclin-dependent kinase inhibitor p16INK4a on lung slides from WT (I) and p66 Shc−/− (J) mice after 5 months of CS exposure. (A-J): Scale bars = 40 μm.
    Dna Fragmentation, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Smoking p66Shc Knocked Out Mice Develop Respiratory Bronchiolitis with Fibrosis but Not Emphysema"

    Article Title: Smoking p66Shc Knocked Out Mice Develop Respiratory Bronchiolitis with Fibrosis but Not Emphysema

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0119797

    Lung slides from WT and p66 Shc−/− mice scored at 5 months for active caspase-3, TUNEL and p16INK4a. Immunohistochemical evaluation of active caspase-3 on lungs of air control (A) and smoking WT (B) mice at 5 months. Note in (B) a positive staining on alveolar macrophages and epithelial lung cells. The positivity on lung epithelial cells is more evident at higher magnification (see the arrowheads in the inset). Active caspase-3 staining on tissue slides from air-control (C) and smoking p66 Shc−/− mice (D). DNA strand-break extremities labeling (TUNEL) on lung tissues from air-control (E) and smoking (F) WT at 5 months. In (G) and (H) TUNEL labeling on air-control and smoking p66 Shc−/− mice, respectively. Immunohistochemical staining for the cyclin-dependent kinase inhibitor p16INK4a on lung slides from WT (I) and p66 Shc−/− (J) mice after 5 months of CS exposure. (A-J): Scale bars = 40 μm.
    Figure Legend Snippet: Lung slides from WT and p66 Shc−/− mice scored at 5 months for active caspase-3, TUNEL and p16INK4a. Immunohistochemical evaluation of active caspase-3 on lungs of air control (A) and smoking WT (B) mice at 5 months. Note in (B) a positive staining on alveolar macrophages and epithelial lung cells. The positivity on lung epithelial cells is more evident at higher magnification (see the arrowheads in the inset). Active caspase-3 staining on tissue slides from air-control (C) and smoking p66 Shc−/− mice (D). DNA strand-break extremities labeling (TUNEL) on lung tissues from air-control (E) and smoking (F) WT at 5 months. In (G) and (H) TUNEL labeling on air-control and smoking p66 Shc−/− mice, respectively. Immunohistochemical staining for the cyclin-dependent kinase inhibitor p16INK4a on lung slides from WT (I) and p66 Shc−/− (J) mice after 5 months of CS exposure. (A-J): Scale bars = 40 μm.

    Techniques Used: Mouse Assay, TUNEL Assay, Immunohistochemistry, Staining, Labeling

    2) Product Images from "MPP+-induced cytotoxicity in neuroblastoma cells: Antagonism and reversal by guanosine"

    Article Title: MPP+-induced cytotoxicity in neuroblastoma cells: Antagonism and reversal by guanosine

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-007-9073-z

    MPP + induces DNA fragmentation in SH-SY5Y cells. MPP + (10, 100 or 500 μM) was added to SH-SY5Y cells and DNA fragmentation determined after 24, 48 or 72 h by oligonucleosomal ELISA as described in the “ Materials and methods ” section. Data are presented as the percentage of DNA fragmentation relative to a positive control ( n > 3). MPP + (500 μM) increased DNA fragmentation significantly at 48 h (* p
    Figure Legend Snippet: MPP + induces DNA fragmentation in SH-SY5Y cells. MPP + (10, 100 or 500 μM) was added to SH-SY5Y cells and DNA fragmentation determined after 24, 48 or 72 h by oligonucleosomal ELISA as described in the “ Materials and methods ” section. Data are presented as the percentage of DNA fragmentation relative to a positive control ( n > 3). MPP + (500 μM) increased DNA fragmentation significantly at 48 h (* p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control

    Guanosine antagonizes and reverses MPP + -induced DNA fragmentation in SH-SY5Y cells. SH-SY5Y cells were exposed to MPP + (500 μM) for 48 h to induce DNA fragmentation. Some cells were treated with guanosine (100 μM) either for 1 h prior to MPP + ( pre-treatment ), or at the same time as MPP + ( co-treatment ) or 24 or 48 h after MPP + ( post-treatment , see Fig. 1 ). DNA fragmentation was determined after 24, 48 or 72 h of MPP + addition by oligonucleosomal ELISA as described in the “ Materials and methods ” section and it is expressed as a percentage of the positive control ( n > 3). MPP + induced a significant increase in DNA fragmentation at 48 (* p
    Figure Legend Snippet: Guanosine antagonizes and reverses MPP + -induced DNA fragmentation in SH-SY5Y cells. SH-SY5Y cells were exposed to MPP + (500 μM) for 48 h to induce DNA fragmentation. Some cells were treated with guanosine (100 μM) either for 1 h prior to MPP + ( pre-treatment ), or at the same time as MPP + ( co-treatment ) or 24 or 48 h after MPP + ( post-treatment , see Fig. 1 ). DNA fragmentation was determined after 24, 48 or 72 h of MPP + addition by oligonucleosomal ELISA as described in the “ Materials and methods ” section and it is expressed as a percentage of the positive control ( n > 3). MPP + induced a significant increase in DNA fragmentation at 48 (* p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control

    Pre-treatment of SH-SY5Y cells with guanosine decreases MPP + -induced DNA fragmentation. SH-SY5Y cells were exposed to MPP + (500 μM) for 48 h to induce DNA fragmentation. Some cells were pre-treated with guanosine (1–300 μM) for 1 h prior to MPP + . Guanosine and MPP + remained in the cultures for the duration of the experiment. DNA fragmentation was determined after 48 h by oligonucleosomal ELISA as described in the “ Materials and methods ” section and is expressed as the percentage of the positive control ( n > 3). MPP + increased DNA fragmentation significantly at 48 h compared to that of untreated controls (* p
    Figure Legend Snippet: Pre-treatment of SH-SY5Y cells with guanosine decreases MPP + -induced DNA fragmentation. SH-SY5Y cells were exposed to MPP + (500 μM) for 48 h to induce DNA fragmentation. Some cells were pre-treated with guanosine (1–300 μM) for 1 h prior to MPP + . Guanosine and MPP + remained in the cultures for the duration of the experiment. DNA fragmentation was determined after 48 h by oligonucleosomal ELISA as described in the “ Materials and methods ” section and is expressed as the percentage of the positive control ( n > 3). MPP + increased DNA fragmentation significantly at 48 h compared to that of untreated controls (* p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Positive Control

    3) Product Images from "BAICALEIN, A COMPONENT OF SCUTELLARIA BAICALENSIS, INDUCES APOPTOSIS BY MCL-1 DOWN-REGULATION IN HUMAN PANCREATIC CANCER CELLS"

    Article Title: BAICALEIN, A COMPONENT OF SCUTELLARIA BAICALENSIS, INDUCES APOPTOSIS BY MCL-1 DOWN-REGULATION IN HUMAN PANCREATIC CANCER CELLS

    Journal: Biochimica et biophysica acta

    doi: 10.1016/j.bbamcr.2011.05.003

    Role of Mcl-1 in baicalein-induced apoptosis. Cells were transiently transfected with specific siRNA against Bcl-2, Bcl-xL, Mcl-1, or control siRNA for 24 h. Anti-apoptotic Bcl-2 family protein expression (A, lower panel) and DNA fragmentation (A, upper
    Figure Legend Snippet: Role of Mcl-1 in baicalein-induced apoptosis. Cells were transiently transfected with specific siRNA against Bcl-2, Bcl-xL, Mcl-1, or control siRNA for 24 h. Anti-apoptotic Bcl-2 family protein expression (A, lower panel) and DNA fragmentation (A, upper

    Techniques Used: Transfection, Expressing

    The effect of baicalein on cell apoptosis. A, cells were seeded in 6-well plates and treated with baicalein for 24 h, after which DNA fragmentation was measured. Increase in apoptosis (DNA fragmentation) is presented as enrichment factor. * p
    Figure Legend Snippet: The effect of baicalein on cell apoptosis. A, cells were seeded in 6-well plates and treated with baicalein for 24 h, after which DNA fragmentation was measured. Increase in apoptosis (DNA fragmentation) is presented as enrichment factor. * p

    Techniques Used:

    4) Product Images from "Sperm chromatin and DNA integrity, methyltransferase mRNA levels, and global DNA methylation in oligoasthenoteratozoospermia"

    Article Title: Sperm chromatin and DNA integrity, methyltransferase mRNA levels, and global DNA methylation in oligoasthenoteratozoospermia

    Journal: Clinical and Experimental Reproductive Medicine

    doi: 10.5653/cerm.2018.45.1.17

    TUNEL assay for the detection of sperm DNA fragmentation. Under fluorescent microscopy, normal DNA is seen as light green (TUNEL−) and damaged DNA is seen as bright green (TUNEL+) (×100 eyepiece magnification). TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.
    Figure Legend Snippet: TUNEL assay for the detection of sperm DNA fragmentation. Under fluorescent microscopy, normal DNA is seen as light green (TUNEL−) and damaged DNA is seen as bright green (TUNEL+) (×100 eyepiece magnification). TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

    Techniques Used: TUNEL Assay, Microscopy

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Interferon-? Reduces the Proliferation of Primed Human Renal Tubular Cells
    Article Snippet: MTT Assay The viable cell number was determined by incubating cell cultures with 0.5 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) for 4 h. Then, 10% sodium dodecyl sulfate in 0.01 M HCl was added 1:1 (vol/vol) and left overnight at 37°C. .. DNA Fragmentation DNA fragmentation was determined by ELISA with the commercial kit Cell Death Detection Plus (Roche Diagnostics, Barcelona, Spain) according to the manufacturer's instructions. ..

    Article Title: Ligands of the peripheral benzodiazepine receptor induce apoptosis and cell cycle arrest in oesophageal cancer cells: involvement of the p38MAPK signalling pathway
    Article Snippet: The cleavage of DEVD-AMC was measured with a VersaFluor fluorometer (excitation: 360 nm emission: 460 nm) from Biorad, Munich, Germany ( ). .. DNA fragmentation DNA fragmentation was determined by Cell Death Detection ELISA (Roche Molecular Biochemicals) according to the manufacturer's instructions ( ). .. Briefly, after incubation with the indicated compounds, cells were lysed in incubation buffer.

    Article Title: NADPH Oxidase versus Mitochondria-Derived ROS in Glucose-Induced Apoptosis of Pericytes in Early Diabetic Retinopathy
    Article Snippet: .. DNA Fragmentation DNA fragmentation was quantified using the Cell Death Detection ELISA plus (1774425; Roche, Hertfordshire, UK). ..

    DNA Fragmentation Assay:

    Article Title: Induction of Cell Death in the Human Acute Lymphoblastic Leukemia Cell Line Reh by Infection with Rotavirus Isolate Wt1-5
    Article Snippet: The images were acquired using NIS-Elements Advanced Research software (Nikon Instruments, Tokyo, Japan) and were analyzed with the ImageJ 1.44p Java 1.6.0_20 (32-bit) software. .. DNA Fragmentation Assay DNA fragmentation was assessed using the Apoptotic DNA Ladder Kit (Roche® Cat. No. 11 835 246 001, Branchburg, NJ, USA). .. Rotavirus-infected cells (2.6 × 106 cells/mL) were harvested at 12 and 24 h.p.i. and resuspended in PBS (200 µL) containing 0.5 mM PMSF and mixed with lysis buffer (200 µL) (6 M guanidine-HCl, 10 M urea, 10 mM Tris-HCl, pH 4.4, 20% Triton X-100).

    Article Title: Ionizing Radiation Regulates Vascular Endothelial Growth Factor-A Transcription in Cultured Human Vascular Endothelial Cells Via the PERK/eIF2α/ATF4 Pathway.
    Article Snippet: .. The delivery of high-dose hypofractionated radiation to a tumor induces vascular damage, but little is known about the responses of vascular endothelial cells to high-dose radiation. ..

    Article Title: Para-Toluenesulfonamide Induces Anti-tumor Activity Through Akt-Dependent and -Independent mTOR/p70S6K Pathway: Roles of Lipid Raft and Cholesterol Contents
    Article Snippet: After incubation for 6 h at 37°C, cells were washed and incubated in 10% FBS-containing RPMI-1640 medium for 48 h. The cells were treated with or without the compound. .. DNA Fragmentation Assay DNA fragmentation was determined using Cell Death Detection ELISAplus kit (Roche, Mannheim, Germany). .. The assay was based on quantitative in vitro determination of cytoplasmic histone-related DNA fragments (mono- and oligonucleosomes).

    End Labeling:

    Article Title: Preclinical Assessment of the Treatment of Second-Stage African Trypanosomiasis with Cordycepin and Deoxycoformycin
    Article Snippet: DNA content Briefly, following a 30 min permeabilization of trypanosomes in 10 mM phosphate buffer containing 6 µM digitonin, nuclei were stained with a 10 µg/ml propidium iodide solution in 10 mM phosphate buffer and kept on ice until measurement by flow cytometry. .. DNA fragmentation DNA fragmentation was assessed by using the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling method (Roche) as described . ..

    In Situ:

    Article Title: Intracellular Acidosis Promotes Mitochondrial Apoptosis Pathway: Role of EMMPRIN Down-regulation via Specific Single-chain Fv Intrabody
    Article Snippet: .. DNA fragmentation DNA fragmentation was carried out using the In Situ Cell Death Detection Kit, TMR red (Roche, Mannheim, Germany), according to the instructions provided by the manufacturer. .. Briefly, after 48 h of transduction with 100 MOI of Ad5/F35-scFv-irrelevant or Ad5/F35-scFv-M6-1B9 , the cells were fixed with the fixative solution (4% Paraformaldehyde in PBS pH 7.4) for 1 h at room temperature.

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    Roche cytoplasmic histone associated dna fragments
    CpG-1-PTO and nCpG-5-PTO amplify staurosporin-induced caspase 9 activity and cytoplasmic histone-assosiated <t>DNA</t> fragments in HaCaT cells—CpG-1-PTO protects RAW264.7 cells from <t>apoptosis.</t> HaCaT cells were treated with 8 µM CpG-1-PTO or nCpG-5-PTO in the presence of 0.1, 0.5 and 1 µM staurosporin (STS). ( A ) After 24 h caspase 9 activity was assessed using a commercial assay as described under ‘Materials and Methods’ section. Each bar represents the mean of four parallel experiments; the standard deviations are indicated (* P
    Cytoplasmic Histone Associated Dna Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche cellular dna fragmentation elisa
    Effect of PCB3 (300 nM) and its metabolites on a) <t>DNA</t> fragmentation by as determined <t>ELISA</t> b) DNA fragmentation as determined by TUNEL assay c) caspase-8 activity and d) caspase-9 activity after 27 hrs of growth in medium deprived of estrogen by treatment
    Cellular Dna Fragmentation Elisa, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche apoptotic dna ladder kit
    Confirmation of the microscopic observation of apoptosis by the <t>DNA</t> fragmentation assay. Gel-like image showing positive controls. Ladder, Agilent DNA 1000 Kit; Positive Control, Positive control (U937 cells, <t>Apoptotic</t> DNA Ladder Kit); D-0, control for non-irradiated HPBL; D-0.5 through D-4.0, a display of human DNA from 5 donors irradiated with different doses: 0.5, 1.0, 2.0, 3.0 and 4.0-Gy of protons, respectively (Bioanalyzer 2100 Agilent, USA). With increasing doses of radiation there was a qualitative increase in DNA fragmentation, indicative apoptosis as well necrosis, confirming fluorescence microscopy.
    Apoptotic Dna Ladder Kit, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/apoptotic dna ladder kit/product/Roche
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    Roche cardiomyocytes
    Cell death and oxidative stress analyses for primary <t>cardiomyocytes.</t> (A) Apoptosis was detected by TUNEL assay. Representative photomicrographs of fluorescence are shown. Green nuclei represented apoptosis. All of nuclei were stained with DAPI (blue). After merging, blue-green nuclei represented apoptosis. Arrows represented the blue nuclei that were not apoptosis (Magnification of 100×). Scale bar = 500 μm. (B) Representative photomicrographs of DAB staining are shown. Brown nuclei (arrows) represented apoptosis (magnification of 200×). Scale bar = 50μm. Inset: Magnified region illustrating non-apoptotic cardiomyocytes (blue nucleus) and apoptotic ones (brown nucleus). (C) TUNEL staining was quantified as the percentage of apoptotic cardiomyocytes (brown nuclei) of the whole section area ( n = 3). (D) MDA levels in primary cardiomyocytes are shown ( n = 3). (E,F) Cell death assay for primary cardiomyocytes ( n = 5). Representative photomicrographs of trypan blue staining are shown. Blue cells (arrows) represented cell death (magnification of 100×). Scale bar = 500 μm. Inset: Magnified region illustrating dead cardiomyocytes (blue staining). The columns indicate the mean, and the bars indicate the SD. ∗ p
    Cardiomyocytes, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CpG-1-PTO and nCpG-5-PTO amplify staurosporin-induced caspase 9 activity and cytoplasmic histone-assosiated DNA fragments in HaCaT cells—CpG-1-PTO protects RAW264.7 cells from apoptosis. HaCaT cells were treated with 8 µM CpG-1-PTO or nCpG-5-PTO in the presence of 0.1, 0.5 and 1 µM staurosporin (STS). ( A ) After 24 h caspase 9 activity was assessed using a commercial assay as described under ‘Materials and Methods’ section. Each bar represents the mean of four parallel experiments; the standard deviations are indicated (* P

    Journal: Nucleic Acids Research

    Article Title: Oligonucleotides suppress PKB/Akt and act as superinductors of apoptosis in human keratinocytes

    doi: 10.1093/nar/gkp252

    Figure Lengend Snippet: CpG-1-PTO and nCpG-5-PTO amplify staurosporin-induced caspase 9 activity and cytoplasmic histone-assosiated DNA fragments in HaCaT cells—CpG-1-PTO protects RAW264.7 cells from apoptosis. HaCaT cells were treated with 8 µM CpG-1-PTO or nCpG-5-PTO in the presence of 0.1, 0.5 and 1 µM staurosporin (STS). ( A ) After 24 h caspase 9 activity was assessed using a commercial assay as described under ‘Materials and Methods’ section. Each bar represents the mean of four parallel experiments; the standard deviations are indicated (* P

    Article Snippet: Histone-associated DNA fragments Apoptosis was quantified on the basis of cytoplasmic histone-associated DNA fragments using the Cell Death Detection ELISA (Roche, Mannheim, Germany) according to the manufacturer's manual.

    Techniques: Activity Assay

    Effect of PCB3 (300 nM) and its metabolites on a) DNA fragmentation by as determined ELISA b) DNA fragmentation as determined by TUNEL assay c) caspase-8 activity and d) caspase-9 activity after 27 hrs of growth in medium deprived of estrogen by treatment

    Journal: Environment international

    Article Title: Induction of cytochrome P450 1A1 in MCF-7 human breast cancer cells by 4-chlorobiphenyl (PCB3) and the effects of its hydroxylated metabolites on cellular apoptosis

    doi: 10.1016/j.envint.2009.06.012

    Figure Lengend Snippet: Effect of PCB3 (300 nM) and its metabolites on a) DNA fragmentation by as determined ELISA b) DNA fragmentation as determined by TUNEL assay c) caspase-8 activity and d) caspase-9 activity after 27 hrs of growth in medium deprived of estrogen by treatment

    Article Snippet: DNA fragmentation was determined using the ‘Cellular DNA Fragmentation ELISA’ kit (Roche Molecular Biochemicals).

    Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay, Activity Assay

    Effect of PCB3 and its metabolites (300 nM) on: a) DNA fragmentation as determined by ELISA, b) DNA fragmentation as determined by TUNEL assay, c) caspase-8 activity, and d) caspase-9 activity after 27 hr of growth in medium supplemented with 5% FBS.

    Journal: Environment international

    Article Title: Induction of cytochrome P450 1A1 in MCF-7 human breast cancer cells by 4-chlorobiphenyl (PCB3) and the effects of its hydroxylated metabolites on cellular apoptosis

    doi: 10.1016/j.envint.2009.06.012

    Figure Lengend Snippet: Effect of PCB3 and its metabolites (300 nM) on: a) DNA fragmentation as determined by ELISA, b) DNA fragmentation as determined by TUNEL assay, c) caspase-8 activity, and d) caspase-9 activity after 27 hr of growth in medium supplemented with 5% FBS.

    Article Snippet: DNA fragmentation was determined using the ‘Cellular DNA Fragmentation ELISA’ kit (Roche Molecular Biochemicals).

    Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay, Activity Assay

    Confirmation of the microscopic observation of apoptosis by the DNA fragmentation assay. Gel-like image showing positive controls. Ladder, Agilent DNA 1000 Kit; Positive Control, Positive control (U937 cells, Apoptotic DNA Ladder Kit); D-0, control for non-irradiated HPBL; D-0.5 through D-4.0, a display of human DNA from 5 donors irradiated with different doses: 0.5, 1.0, 2.0, 3.0 and 4.0-Gy of protons, respectively (Bioanalyzer 2100 Agilent, USA). With increasing doses of radiation there was a qualitative increase in DNA fragmentation, indicative apoptosis as well necrosis, confirming fluorescence microscopy.

    Journal: Clinical and Translational Radiation Oncology

    Article Title: Do protons and X-rays induce cell-killing in human peripheral blood lymphocytes by different mechanisms?

    doi: 10.1016/j.ctro.2018.01.004

    Figure Lengend Snippet: Confirmation of the microscopic observation of apoptosis by the DNA fragmentation assay. Gel-like image showing positive controls. Ladder, Agilent DNA 1000 Kit; Positive Control, Positive control (U937 cells, Apoptotic DNA Ladder Kit); D-0, control for non-irradiated HPBL; D-0.5 through D-4.0, a display of human DNA from 5 donors irradiated with different doses: 0.5, 1.0, 2.0, 3.0 and 4.0-Gy of protons, respectively (Bioanalyzer 2100 Agilent, USA). With increasing doses of radiation there was a qualitative increase in DNA fragmentation, indicative apoptosis as well necrosis, confirming fluorescence microscopy.

    Article Snippet: Confirmation of apoptosis and necrosis DNA extraction : The DNA from the HPBL was isolated with a commercial DNA isolation kit as a part of Apoptotic DNA Ladder Kit (Roche, Germany) according to the manufacturer’s protocol.

    Techniques: DNA Fragmentation Assay, Positive Control, Irradiation, Fluorescence, Microscopy

    Cell death and oxidative stress analyses for primary cardiomyocytes. (A) Apoptosis was detected by TUNEL assay. Representative photomicrographs of fluorescence are shown. Green nuclei represented apoptosis. All of nuclei were stained with DAPI (blue). After merging, blue-green nuclei represented apoptosis. Arrows represented the blue nuclei that were not apoptosis (Magnification of 100×). Scale bar = 500 μm. (B) Representative photomicrographs of DAB staining are shown. Brown nuclei (arrows) represented apoptosis (magnification of 200×). Scale bar = 50μm. Inset: Magnified region illustrating non-apoptotic cardiomyocytes (blue nucleus) and apoptotic ones (brown nucleus). (C) TUNEL staining was quantified as the percentage of apoptotic cardiomyocytes (brown nuclei) of the whole section area ( n = 3). (D) MDA levels in primary cardiomyocytes are shown ( n = 3). (E,F) Cell death assay for primary cardiomyocytes ( n = 5). Representative photomicrographs of trypan blue staining are shown. Blue cells (arrows) represented cell death (magnification of 100×). Scale bar = 500 μm. Inset: Magnified region illustrating dead cardiomyocytes (blue staining). The columns indicate the mean, and the bars indicate the SD. ∗ p

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Cell death and oxidative stress analyses for primary cardiomyocytes. (A) Apoptosis was detected by TUNEL assay. Representative photomicrographs of fluorescence are shown. Green nuclei represented apoptosis. All of nuclei were stained with DAPI (blue). After merging, blue-green nuclei represented apoptosis. Arrows represented the blue nuclei that were not apoptosis (Magnification of 100×). Scale bar = 500 μm. (B) Representative photomicrographs of DAB staining are shown. Brown nuclei (arrows) represented apoptosis (magnification of 200×). Scale bar = 50μm. Inset: Magnified region illustrating non-apoptotic cardiomyocytes (blue nucleus) and apoptotic ones (brown nucleus). (C) TUNEL staining was quantified as the percentage of apoptotic cardiomyocytes (brown nuclei) of the whole section area ( n = 3). (D) MDA levels in primary cardiomyocytes are shown ( n = 3). (E,F) Cell death assay for primary cardiomyocytes ( n = 5). Representative photomicrographs of trypan blue staining are shown. Blue cells (arrows) represented cell death (magnification of 100×). Scale bar = 500 μm. Inset: Magnified region illustrating dead cardiomyocytes (blue staining). The columns indicate the mean, and the bars indicate the SD. ∗ p

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: TUNEL Assay, Fluorescence, Staining, Multiple Displacement Amplification

    Schematic depicting the mechanisms by which molecular hydrogen mediates apoptosis. H 2 eliminated oxygen free radicals which led p53 phosphorylation. As a transcription factor, phosphorylated p53 increased expression of Bax which increased expression of caspase-3. Cleaved-caspase-3 executed apoptosis of cardiomyocytes. Hence, apoptosis was suppressed by inhibition of p53 in response to molecular hydrogen.

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Schematic depicting the mechanisms by which molecular hydrogen mediates apoptosis. H 2 eliminated oxygen free radicals which led p53 phosphorylation. As a transcription factor, phosphorylated p53 increased expression of Bax which increased expression of caspase-3. Cleaved-caspase-3 executed apoptosis of cardiomyocytes. Hence, apoptosis was suppressed by inhibition of p53 in response to molecular hydrogen.

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: Expressing, Inhibition

    Effects of H 2 on apoptosis and cardiac p53 protein expression in CHF. (A) Apoptosis was detected by TUNEL assay (brown nuclei, arrows). Representative photomicrographs are shown (magnification of 200×). Scale bar = 50 μm. Inset: Magnified region illustrating non-apoptotic cardiomyocytes (blue nucleus) and apoptotic ones (brown nucleus). (B) Nuclei of cardiomyocyte was evaluated by observation under electronic microscopy (magnification of 10,000×). Scale bar = 2 μm. (C) TUNEL staining was quantified as the percentage of apoptotic cardiomyocytes of the whole section area ( n = 8–9). (D,E) Confirmation by Western blot analyses for the expressions of cardiac Bax and cleaved-caspase-3. The ratios of Bax and bcl-2 are shown ( n = 8–10). (F) Representative photomicrographs of immunohistochemistry about p53 (brown nuclei, arrows) are shown (magnification of 200×). Scale bar = 50 μm. Inset: Magnified region illustrating p53 negative cardiomyocytes (blue nucleus) and positive ones (brown nucleus). (G) P53 was determined by measuring the positive area and was expressed as a percentage of the whole section area ( n = 8–9). (H,I) Confirmation by Western blot analyses for the expression of cardiac p53 and phospho-p53. The ratios of phospho-p53 and p53 are shown ( n = 10). Representative photographs are shown. GAPDH was used as a loading control. Expressions of cleaved-caspase-3, Bax, p53 and phospho-p53 were, respectively, quantified as folds of GAPDH. The columns indicate the mean, and the bars indicate the SD (TUNEL and p53 staining) or SE (Western blotting). ∗ p

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Effects of H 2 on apoptosis and cardiac p53 protein expression in CHF. (A) Apoptosis was detected by TUNEL assay (brown nuclei, arrows). Representative photomicrographs are shown (magnification of 200×). Scale bar = 50 μm. Inset: Magnified region illustrating non-apoptotic cardiomyocytes (blue nucleus) and apoptotic ones (brown nucleus). (B) Nuclei of cardiomyocyte was evaluated by observation under electronic microscopy (magnification of 10,000×). Scale bar = 2 μm. (C) TUNEL staining was quantified as the percentage of apoptotic cardiomyocytes of the whole section area ( n = 8–9). (D,E) Confirmation by Western blot analyses for the expressions of cardiac Bax and cleaved-caspase-3. The ratios of Bax and bcl-2 are shown ( n = 8–10). (F) Representative photomicrographs of immunohistochemistry about p53 (brown nuclei, arrows) are shown (magnification of 200×). Scale bar = 50 μm. Inset: Magnified region illustrating p53 negative cardiomyocytes (blue nucleus) and positive ones (brown nucleus). (G) P53 was determined by measuring the positive area and was expressed as a percentage of the whole section area ( n = 8–9). (H,I) Confirmation by Western blot analyses for the expression of cardiac p53 and phospho-p53. The ratios of phospho-p53 and p53 are shown ( n = 10). Representative photographs are shown. GAPDH was used as a loading control. Expressions of cleaved-caspase-3, Bax, p53 and phospho-p53 were, respectively, quantified as folds of GAPDH. The columns indicate the mean, and the bars indicate the SD (TUNEL and p53 staining) or SE (Western blotting). ∗ p

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: Expressing, TUNEL Assay, Microscopy, Staining, Western Blot, Immunohistochemistry

    Left ventricular remodeling size and oxidative stress damage in myocardial tissue after inhalation of H 2 and air. (A) Representative photomicrographs of Masson’s trichrome-stained heart sections are shown (magnification of 200×). Blue area represents fibrosis. Scale bars = 50 μm. (B) Representative photomicrographs of immunohistochemistry about 8-OHdG (brown nuclei, arrows) are shown (magnification of 200×). Scale bars = 50 μm. Inset: Magnified region illustrating 8-OHdG negative cardiomyocytes (blue nucleus) and positive ones (brown nucleus). (C) Left ventricular remodeling area was determined by measuring the area of fibrosis and was expressed as a percentage of the whole LV area ( n = 7–9). (D) 8-OHdG was determined by measuring the positive area of brown nucleus and was expressed as a percentage of the whole section area ( n = 8–10). (E) MDA levels in myocardial tissue are shown ( n = 12–14). The columns indicate the mean, and the bars indicate the SD (LVEDP, LVESP, and MDA) or SE (+dp/dt, –dp/dt, Masson, and 8-OHdG). ∗ p

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Left ventricular remodeling size and oxidative stress damage in myocardial tissue after inhalation of H 2 and air. (A) Representative photomicrographs of Masson’s trichrome-stained heart sections are shown (magnification of 200×). Blue area represents fibrosis. Scale bars = 50 μm. (B) Representative photomicrographs of immunohistochemistry about 8-OHdG (brown nuclei, arrows) are shown (magnification of 200×). Scale bars = 50 μm. Inset: Magnified region illustrating 8-OHdG negative cardiomyocytes (blue nucleus) and positive ones (brown nucleus). (C) Left ventricular remodeling area was determined by measuring the area of fibrosis and was expressed as a percentage of the whole LV area ( n = 7–9). (D) 8-OHdG was determined by measuring the positive area of brown nucleus and was expressed as a percentage of the whole section area ( n = 8–10). (E) MDA levels in myocardial tissue are shown ( n = 12–14). The columns indicate the mean, and the bars indicate the SD (LVEDP, LVESP, and MDA) or SE (+dp/dt, –dp/dt, Masson, and 8-OHdG). ∗ p

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: Staining, Immunohistochemistry, Multiple Displacement Amplification

    Analysis for effects of H 2 on the expression of p53 in H9c2 cells. (A–D) Overexpression of p53 in H9c2 cells. In the groups of P53 and pLVX, H9c2 cells were infected with p53-lentivirus or pLVX and were selected for 72 h. Then the cells were treated with 10 μM VK 3 and 75% H 2 for 8 h. In the groups of DMSO, VK 3 and VK 3 +H 2 , the cells were treated as the primary cardiomyocytes. (E–H) Inhibition of p53 in H9c2 cells. H9c2 cells were treated with 10 μM PFTα or equivoluminal DMSO for 16 h (DMEM group and PFTα), and then treated with 10 μM VK3 for 8 h (VK3 group and PFTα+VK 3 group). Western blot analysis was performed on 100 g cell lysates with antibodies against p53, Bax, or cleaved caspase-3. GAPDH was used as a loading control. Expressions of p53, Bax, and cleaved caspase-3 were, respectively, quantified as folds of GAPDH ( n = 3–4). The columns indicate the mean, and the bars indicate the SE. ∗ p

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Analysis for effects of H 2 on the expression of p53 in H9c2 cells. (A–D) Overexpression of p53 in H9c2 cells. In the groups of P53 and pLVX, H9c2 cells were infected with p53-lentivirus or pLVX and were selected for 72 h. Then the cells were treated with 10 μM VK 3 and 75% H 2 for 8 h. In the groups of DMSO, VK 3 and VK 3 +H 2 , the cells were treated as the primary cardiomyocytes. (E–H) Inhibition of p53 in H9c2 cells. H9c2 cells were treated with 10 μM PFTα or equivoluminal DMSO for 16 h (DMEM group and PFTα), and then treated with 10 μM VK3 for 8 h (VK3 group and PFTα+VK 3 group). Western blot analysis was performed on 100 g cell lysates with antibodies against p53, Bax, or cleaved caspase-3. GAPDH was used as a loading control. Expressions of p53, Bax, and cleaved caspase-3 were, respectively, quantified as folds of GAPDH ( n = 3–4). The columns indicate the mean, and the bars indicate the SE. ∗ p

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: Expressing, Over Expression, Infection, Inhibition, Western Blot

    Analysis of effects of H 2 on primary cardiomyocytes with oxidative stress. (A,B) Identification of primary cardiomyocytes. Representative photomicrographs of fluorescence are shown. Myofilaments were marked with the green fluorescence, representing the expression of TnT ( B , arrows) which was negative in fibroblasts ( A , arrows) (magnification of 100×). Scale bar = 500 μm. (C–E) Confirmation by Western blot analyses for the expression of cardiac cleaved caspase-3, phospho-p53, p53, and Bax in primary cardiomyocytes. Representative photographs are shown. GAPDH was used as a loading control. Expressions of cleaved-caspase-3, phospho-p53, p53 and Bax were, respectively, quantified as folds of GAPDH ( n = 3–5). The columns indicate the mean, and the bars indicate the SD (MDA and cell viability) or SE (Western blotting). ∗ p

    Journal: Frontiers in Physiology

    Article Title: Inhalation of Hydrogen Attenuates Progression of Chronic Heart Failure via Suppression of Oxidative Stress and P53 Related to Apoptosis Pathway in Rats

    doi: 10.3389/fphys.2018.01026

    Figure Lengend Snippet: Analysis of effects of H 2 on primary cardiomyocytes with oxidative stress. (A,B) Identification of primary cardiomyocytes. Representative photomicrographs of fluorescence are shown. Myofilaments were marked with the green fluorescence, representing the expression of TnT ( B , arrows) which was negative in fibroblasts ( A , arrows) (magnification of 100×). Scale bar = 500 μm. (C–E) Confirmation by Western blot analyses for the expression of cardiac cleaved caspase-3, phospho-p53, p53, and Bax in primary cardiomyocytes. Representative photographs are shown. GAPDH was used as a loading control. Expressions of cleaved-caspase-3, phospho-p53, p53 and Bax were, respectively, quantified as folds of GAPDH ( n = 3–5). The columns indicate the mean, and the bars indicate the SD (MDA and cell viability) or SE (Western blotting). ∗ p

    Article Snippet: TUNEL Staining The TUNEL assay was carried out to label the 3′-end of fragmented DNA in tissue slices or cardiomyocytes in accordance with the manufacturer’s directions (Roche, Switzerland) and stained with DAB kit (ZLI-9018, ZSGB-BIO, China).

    Techniques: Fluorescence, Expressing, Western Blot, Multiple Displacement Amplification