ethidium bromide Search Results


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  • 94
    Thermo Fisher ethidium bromide
    2.3% native agarose gel in 0.5X TBE buffer, showing a representative library prep from wild-type Okazaki fragments. The gel is stained with <t>Ethidium</t> Bromide. Monosome- and disome-sized fragments with ligated sequencing-adaptors can be seen along with
    Ethidium Bromide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 10037 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore ethidium bromide monoazide
    2.3% native agarose gel in 0.5X TBE buffer, showing a representative library prep from wild-type Okazaki fragments. The gel is stained with <t>Ethidium</t> Bromide. Monosome- and disome-sized fragments with ligated sequencing-adaptors can be seen along with
    Ethidium Bromide Monoazide, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam ethidium bromide uptake
    Evaluation of astrocytic GJIC permeability and hemichannel activity after OGD/R injury with Gap19 or Gap26. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and cocultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R injury decreased the degree of astrocytic coupling; Gap26 further inhibited cellular coupling. c Representative images depicting <t>ethidium</t> uptake via hemichannels in the four groups. d OGD/R injury increased astrocytic ethidium uptake, while Gap19 and Gap26 achieved similar attenuation of hemichannel opening. e The supernatant ATP concentration was strongly elevated in the OGD/R group astrocytes, but Gap19 and Gap26 reversed this effect. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p
    Ethidium Bromide Uptake, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Bio-Rad ethidium bromide
    The ovarian content of the mRNAs encoding GDF-9 and its presumed receptor, BMPRII, and KL and its c-kit receptor is not altered in trkB −/− mice. (A and B) Representative RealTime PCR amplification profiles of GDF-9 mRNA (A) and KL mRNA (B) from the ovary of trkB −/− mice compared with that of wild-type littermates. The insets depict the standard curves use to estimate the amount of each mRNA present in the two groups. The curves were generated using serial dilutions of reverse transcription reactions derived from one μg of total RNA from 7-day-old mouse ovaries (equivalent to 10 −1 – 10 −8 ) using an ABI Prism 7700 sequence Detection System (Perkin Elmer Applied Biosystems), in a total volume of 10 μl, containing 2 μl of reverse transcription reaction, 5 μl of TaqMan Universal PCR Master Mix (Perkin Elmer Applied Biosystems), and 3 μl primer mix (250 nM of each gene-specific and ribosomal fluorescent probes and 300 nM of each gene-specific unlabeled primer plus 80 nM of each unlabeled ribosomal primer). (C and D) Ovarian content of GDF-9 (C) and KL (D) mRNA detected by RealTime PCR in wild-type and trkB −/− 7-day-old mice. (E and F) Representative <t>ethidium</t> bromide-stained gels showing BMPRII (E) and c-kit (F) PCR products amplified from total ovarian RNA using the Relative Quantitative PCR conditions described in Materials and methods. (G and H) Relative content of BMPRII (G) and c-kit (H) mRNAs estimated by Relative Quantitative PCR. Values are shown as ratios between the gene-specific signal and the signal provided by 18S ribosomal RNA coamplified with each gene of interest. Bars are mean ± SEM and numbers on top of bars are number of mice per group. Because the KL and BMPRII mRNAs are expressed in granulosa cells, the values obtained in trkB −/− mice were corrected to take into consideration the reduction in granulosa cell number observed in these animals.
    Ethidium Bromide, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 19715 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BioVision ethidium bromide
    The ovarian content of the mRNAs encoding GDF-9 and its presumed receptor, BMPRII, and KL and its c-kit receptor is not altered in trkB −/− mice. (A and B) Representative RealTime PCR amplification profiles of GDF-9 mRNA (A) and KL mRNA (B) from the ovary of trkB −/− mice compared with that of wild-type littermates. The insets depict the standard curves use to estimate the amount of each mRNA present in the two groups. The curves were generated using serial dilutions of reverse transcription reactions derived from one μg of total RNA from 7-day-old mouse ovaries (equivalent to 10 −1 – 10 −8 ) using an ABI Prism 7700 sequence Detection System (Perkin Elmer Applied Biosystems), in a total volume of 10 μl, containing 2 μl of reverse transcription reaction, 5 μl of TaqMan Universal PCR Master Mix (Perkin Elmer Applied Biosystems), and 3 μl primer mix (250 nM of each gene-specific and ribosomal fluorescent probes and 300 nM of each gene-specific unlabeled primer plus 80 nM of each unlabeled ribosomal primer). (C and D) Ovarian content of GDF-9 (C) and KL (D) mRNA detected by RealTime PCR in wild-type and trkB −/− 7-day-old mice. (E and F) Representative <t>ethidium</t> bromide-stained gels showing BMPRII (E) and c-kit (F) PCR products amplified from total ovarian RNA using the Relative Quantitative PCR conditions described in Materials and methods. (G and H) Relative content of BMPRII (G) and c-kit (H) mRNAs estimated by Relative Quantitative PCR. Values are shown as ratios between the gene-specific signal and the signal provided by 18S ribosomal RNA coamplified with each gene of interest. Bars are mean ± SEM and numbers on top of bars are number of mice per group. Because the KL and BMPRII mRNAs are expressed in granulosa cells, the values obtained in trkB −/− mice were corrected to take into consideration the reduction in granulosa cell number observed in these animals.
    Ethidium Bromide, supplied by BioVision, used in various techniques. Bioz Stars score: 93/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Promega ethidium
    LmrP mediates cellular sensitivity to Hoechst 33342 but resistance to <t>ethidium.</t> Growth rate of LmrP-expressing lactococcal cells and cells without LmrP expression (control) in the presence of up to 8.4 μM Hoechst 33342 (A), or up to 24 μM ethidium (B) was determined relative to the maximum growth rate in the absence of drug. The error bars for some of the data points were too small to be displayed, and are hidden behind the data point symbols.
    Ethidium, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Amresco ethidium bromide
    NAI or NAI-imine increase P2X7-mediated IL-1 β release but not <t>ethidium</t> + uptake. LPS-primed J774 cells in physiological medium were preincubated in the presence of DMSO, or 1 μ M NAI or 1 μ M NAI-imine for 30 min, and then in the absence or presence of 3 mM ATP for ((a) and (b)) 20 min or (c) 5 min in the presence of 25 μ M ethidium bromide. (a) The IL-1 β concentration in cell-free supernatants was measured using an ELISA. (b) The LDH activity in cell-free supernatants and cell lysates was measured using a cytotoxicity kit, and results are expressed as a percentage of maximal LDH release from lysed cells. (c) Ethidium + uptake was measured by flow cytometry and is expressed as mean fluorescence intensity (MFI). ((a) to (c)) Results are mean ± SD ( n = 3); ∗ P
    Ethidium Bromide, supplied by Amresco, used in various techniques. Bioz Stars score: 92/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck & Co ethidium bromide
    NAI or NAI-imine increase P2X7-mediated IL-1 β release but not <t>ethidium</t> + uptake. LPS-primed J774 cells in physiological medium were preincubated in the presence of DMSO, or 1 μ M NAI or 1 μ M NAI-imine for 30 min, and then in the absence or presence of 3 mM ATP for ((a) and (b)) 20 min or (c) 5 min in the presence of 25 μ M ethidium bromide. (a) The IL-1 β concentration in cell-free supernatants was measured using an ELISA. (b) The LDH activity in cell-free supernatants and cell lysates was measured using a cytotoxicity kit, and results are expressed as a percentage of maximal LDH release from lysed cells. (c) Ethidium + uptake was measured by flow cytometry and is expressed as mean fluorescence intensity (MFI). ((a) to (c)) Results are mean ± SD ( n = 3); ∗ P
    Ethidium Bromide, supplied by Merck & Co, used in various techniques. Bioz Stars score: 92/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck KGaA ethidium bromide
    Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) <t>ethidium</t> bromide; ( C ) YO; ( D ) YOYO.
    Ethidium Bromide, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 286 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioShop ethidium bromide
    Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) <t>ethidium</t> bromide; ( C ) YO; ( D ) YOYO.
    Ethidium Bromide, supplied by BioShop, used in various techniques. Bioz Stars score: 92/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bioneer Corporation ethidium bromide
    Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) <t>ethidium</t> bromide; ( C ) YO; ( D ) YOYO.
    Ethidium Bromide, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 92/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa ethidium bromide
    Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) <t>ethidium</t> bromide; ( C ) YO; ( D ) YOYO.
    Ethidium Bromide, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 475 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    2.3% native agarose gel in 0.5X TBE buffer, showing a representative library prep from wild-type Okazaki fragments. The gel is stained with Ethidium Bromide. Monosome- and disome-sized fragments with ligated sequencing-adaptors can be seen along with

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Detection and sequencing of Okazaki fragments in S. cerevisiae

    doi: 10.1007/978-1-4939-2596-4_10

    Figure Lengend Snippet: 2.3% native agarose gel in 0.5X TBE buffer, showing a representative library prep from wild-type Okazaki fragments. The gel is stained with Ethidium Bromide. Monosome- and disome-sized fragments with ligated sequencing-adaptors can be seen along with

    Article Snippet: YPD (1% yeast extract, 2% peptone: autoclave and add filter-sterilized 40% glucose to a final concentration of 2%) 100mg/ml Doxycycline hydrochloride stock solution in DMSO (if using Dox-repressible CDC9 construct) (Fisher Scientific, Cat #BP2653-5) Shaking incubator 250ml Erlenmeyer flasks SCE: 1M sorbitol, 100mM sodium citrate, 60mM EDTA. pH to 7.0, filter-sterilize and store at 4°C Zymolyase 20T (Sunrise Science Products, N0766391) Lysis buffer: 50mM Tris-HCl pH8.0, 50mM EDTA, 100mM NaCl, 1.5% sarkosyl Proteinase K (Fisher Scientific, cat# BP1700100) Isopropanol 3M Sodium acetate, pH 5.2 Absolute ethanol STE: 100mM NaCl, 10mM Tris-HCl pH8.0, 1mM EDTA Riboshredder RNase blend (Epicentre, cat #RS12500) or RNase A (20mg/ml stock solution) TE: 10mM Tris-HCl pH8.0, 1mM EDTA (Optional) Spin-X centrifuge tubes with 0.45um cellulose acetate filter (Costar, cat#8162) Agarose, 0.5x TAE/0.5x TBE running buffer, and GelRed or ethidium bromide (Invitrogen, Cat#15585-011)

    Techniques: Agarose Gel Electrophoresis, Staining, Sequencing

    Okazaki fragment preparations resolved on 0.7% native agarose gel in 0.5X TBE buffer, stained with Ethidium bromide. Large genomic DNA can be seen above the corresponding 10kb band of 2log ladder, while contaminating RNA typically runs below 100bp as

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Detection and sequencing of Okazaki fragments in S. cerevisiae

    doi: 10.1007/978-1-4939-2596-4_10

    Figure Lengend Snippet: Okazaki fragment preparations resolved on 0.7% native agarose gel in 0.5X TBE buffer, stained with Ethidium bromide. Large genomic DNA can be seen above the corresponding 10kb band of 2log ladder, while contaminating RNA typically runs below 100bp as

    Article Snippet: YPD (1% yeast extract, 2% peptone: autoclave and add filter-sterilized 40% glucose to a final concentration of 2%) 100mg/ml Doxycycline hydrochloride stock solution in DMSO (if using Dox-repressible CDC9 construct) (Fisher Scientific, Cat #BP2653-5) Shaking incubator 250ml Erlenmeyer flasks SCE: 1M sorbitol, 100mM sodium citrate, 60mM EDTA. pH to 7.0, filter-sterilize and store at 4°C Zymolyase 20T (Sunrise Science Products, N0766391) Lysis buffer: 50mM Tris-HCl pH8.0, 50mM EDTA, 100mM NaCl, 1.5% sarkosyl Proteinase K (Fisher Scientific, cat# BP1700100) Isopropanol 3M Sodium acetate, pH 5.2 Absolute ethanol STE: 100mM NaCl, 10mM Tris-HCl pH8.0, 1mM EDTA Riboshredder RNase blend (Epicentre, cat #RS12500) or RNase A (20mg/ml stock solution) TE: 10mM Tris-HCl pH8.0, 1mM EDTA (Optional) Spin-X centrifuge tubes with 0.45um cellulose acetate filter (Costar, cat#8162) Agarose, 0.5x TAE/0.5x TBE running buffer, and GelRed or ethidium bromide (Invitrogen, Cat#15585-011)

    Techniques: Agarose Gel Electrophoresis, Staining

    Effect of Rlip-LNA on apoptosis and DNA fragmentation in MDA-MB-231 and MCF7 cells as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and DNA laddering assays. ( A ) Apoptosis by DNA laddering. After 48 h of treatment with CAS or Rlip-LNA, DNA was extracted and subjected to agarose-gel electrophoresis in 2% agarose with a 1 kB DNA ladder. Ethidium-bromide-stained gels were visualized and photographed under 260 nm UV light. ( B , C ) Effect of Rlip depletion on DNA fragmentation in MDA-MB-231 and MCF7 cells as measured by TUNEL. Cells were transfected with Rlip-LNA or control antisense (CAS) for 24 hours, as described in the Materials and Methods. After Rlip depletion, the apoptotic intensity was determined by flow cytometric TUNEL assay. ( B ) Logarithmic dot plots show the percentage of TUNEL-positive cells in different groups (US = unstained) as measured by flow cytometry. Viable cells were identified by gating on forward and side scatters. ( C ) The overlapped peaks (logarithmic histogram) demonstrate the effects as a whole and are expressed as the fluorescence intensity of the number of counts of the TUNEL-positive cells obtained from the statistical analysis of the fluorescence height and mean value of the x-axis displayed by the software. The fluorescence level for discrimination between apoptotic and nonapoptotic cells was set using the control without TdT (terminal deoxynucleotidyl transferase). Cells above this fluorescence value in the TdT-positive sample were considered apoptotic. Analysis was performed using the BD CSampler software (BD Biosciences). At least 10,000 cells were analyzed per staining. Data were obtained from three independent experiments.

    Journal: Cancers

    Article Title: Rlip Depletion Suppresses Growth of Breast Cancer

    doi: 10.3390/cancers12061446

    Figure Lengend Snippet: Effect of Rlip-LNA on apoptosis and DNA fragmentation in MDA-MB-231 and MCF7 cells as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and DNA laddering assays. ( A ) Apoptosis by DNA laddering. After 48 h of treatment with CAS or Rlip-LNA, DNA was extracted and subjected to agarose-gel electrophoresis in 2% agarose with a 1 kB DNA ladder. Ethidium-bromide-stained gels were visualized and photographed under 260 nm UV light. ( B , C ) Effect of Rlip depletion on DNA fragmentation in MDA-MB-231 and MCF7 cells as measured by TUNEL. Cells were transfected with Rlip-LNA or control antisense (CAS) for 24 hours, as described in the Materials and Methods. After Rlip depletion, the apoptotic intensity was determined by flow cytometric TUNEL assay. ( B ) Logarithmic dot plots show the percentage of TUNEL-positive cells in different groups (US = unstained) as measured by flow cytometry. Viable cells were identified by gating on forward and side scatters. ( C ) The overlapped peaks (logarithmic histogram) demonstrate the effects as a whole and are expressed as the fluorescence intensity of the number of counts of the TUNEL-positive cells obtained from the statistical analysis of the fluorescence height and mean value of the x-axis displayed by the software. The fluorescence level for discrimination between apoptotic and nonapoptotic cells was set using the control without TdT (terminal deoxynucleotidyl transferase). Cells above this fluorescence value in the TdT-positive sample were considered apoptotic. Analysis was performed using the BD CSampler software (BD Biosciences). At least 10,000 cells were analyzed per staining. Data were obtained from three independent experiments.

    Article Snippet: Ethidium-bromide-stained gels were visualized and photographed under 260 nm UV light [ ].

    Techniques: Multiple Displacement Amplification, TUNEL Assay, DNA Laddering, Agarose Gel Electrophoresis, Staining, Transfection, Flow Cytometry, Fluorescence, Software

    Evaluation of astrocytic GJIC permeability and hemichannel activity after OGD/R injury with Gap19 or Gap26. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and cocultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R injury decreased the degree of astrocytic coupling; Gap26 further inhibited cellular coupling. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R injury increased astrocytic ethidium uptake, while Gap19 and Gap26 achieved similar attenuation of hemichannel opening. e The supernatant ATP concentration was strongly elevated in the OGD/R group astrocytes, but Gap19 and Gap26 reversed this effect. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Journal: Journal of Neuroinflammation

    Article Title: Roles of astrocytic connexin-43, hemichannels, and gap junctions in oxygen-glucose deprivation/reperfusion injury induced neuroinflammation and the possible regulatory mechanisms of salvianolic acid B and carbenoxolone

    doi: 10.1186/s12974-018-1127-3

    Figure Lengend Snippet: Evaluation of astrocytic GJIC permeability and hemichannel activity after OGD/R injury with Gap19 or Gap26. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and cocultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R injury decreased the degree of astrocytic coupling; Gap26 further inhibited cellular coupling. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R injury increased astrocytic ethidium uptake, while Gap19 and Gap26 achieved similar attenuation of hemichannel opening. e The supernatant ATP concentration was strongly elevated in the OGD/R group astrocytes, but Gap19 and Gap26 reversed this effect. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Article Snippet: Ethidium bromide uptake For dye uptake experiments, astrocytes cultured were washed and then exposed to 0.5 μM ethidium bromide (EtBr) (Abcam, Cat#ab141391) for 10 min at 37 °C.

    Techniques: Permeability, Activity Assay, Flow Cytometry, Cytometry, Labeling, Concentration Assay

    Evaluation of impact of MCM on astrocytic GJIC permeability and hemichannel activity after OGD/R injury. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and co-cultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R-MCM decreased the degree of astrocytic coupling, but OGD/R-SalB-MCM increased astrocytic intercellular dye transfer. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R-MCM increased astrocytic ethidium uptake, but this effect was reversed in OGD/R-SalB-MCM-treated astrocytes. e OGD/R-MCM increased ATP concentrations in the astrocytic supernatant, but this effect was reversed by OGD/R-SalB-MCM. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Journal: Journal of Neuroinflammation

    Article Title: Roles of astrocytic connexin-43, hemichannels, and gap junctions in oxygen-glucose deprivation/reperfusion injury induced neuroinflammation and the possible regulatory mechanisms of salvianolic acid B and carbenoxolone

    doi: 10.1186/s12974-018-1127-3

    Figure Lengend Snippet: Evaluation of impact of MCM on astrocytic GJIC permeability and hemichannel activity after OGD/R injury. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and co-cultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R-MCM decreased the degree of astrocytic coupling, but OGD/R-SalB-MCM increased astrocytic intercellular dye transfer. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R-MCM increased astrocytic ethidium uptake, but this effect was reversed in OGD/R-SalB-MCM-treated astrocytes. e OGD/R-MCM increased ATP concentrations in the astrocytic supernatant, but this effect was reversed by OGD/R-SalB-MCM. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Article Snippet: Ethidium bromide uptake For dye uptake experiments, astrocytes cultured were washed and then exposed to 0.5 μM ethidium bromide (EtBr) (Abcam, Cat#ab141391) for 10 min at 37 °C.

    Techniques: Permeability, Activity Assay, Flow Cytometry, Cytometry, Labeling, Cell Culture

    Evaluation of astrocytic GJIC permeability and hemichannel activity after OGD/R injury with SalB or CBX. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and cocultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R injury decreased the degree of astrocytic coupling, but SalB reversed this effect. CBX further inhibited cellular coupling. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R injury increased astrocytic ethidium uptake, but SalB and CBX achieved near-significant attenuation of this effect. e The supernatant ATP concentration was strongly elevated in the OGD/R group astrocytes, but SalB and CBX reversed this effect. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Journal: Journal of Neuroinflammation

    Article Title: Roles of astrocytic connexin-43, hemichannels, and gap junctions in oxygen-glucose deprivation/reperfusion injury induced neuroinflammation and the possible regulatory mechanisms of salvianolic acid B and carbenoxolone

    doi: 10.1186/s12974-018-1127-3

    Figure Lengend Snippet: Evaluation of astrocytic GJIC permeability and hemichannel activity after OGD/R injury with SalB or CBX. a For GJIC detection, we measured calcein-AM transfer between “donor cells” and “acceptor cells” with flow cytometry. Shown here is a representative flow cytometry plot of transfer after donor astrocytes were labeled with calcein-AM and cocultured with acceptor astrocytes for 4 h. Grouped dye transfer data are shown in b . OGD/R injury decreased the degree of astrocytic coupling, but SalB reversed this effect. CBX further inhibited cellular coupling. c Representative images depicting ethidium uptake via hemichannels in the four groups. d OGD/R injury increased astrocytic ethidium uptake, but SalB and CBX achieved near-significant attenuation of this effect. e The supernatant ATP concentration was strongly elevated in the OGD/R group astrocytes, but SalB and CBX reversed this effect. We evaluated the statistical significance with ANOVA and Duncan’s multiple comparisons test. * p

    Article Snippet: Ethidium bromide uptake For dye uptake experiments, astrocytes cultured were washed and then exposed to 0.5 μM ethidium bromide (EtBr) (Abcam, Cat#ab141391) for 10 min at 37 °C.

    Techniques: Permeability, Activity Assay, Flow Cytometry, Cytometry, Labeling, Concentration Assay

    The ovarian content of the mRNAs encoding GDF-9 and its presumed receptor, BMPRII, and KL and its c-kit receptor is not altered in trkB −/− mice. (A and B) Representative RealTime PCR amplification profiles of GDF-9 mRNA (A) and KL mRNA (B) from the ovary of trkB −/− mice compared with that of wild-type littermates. The insets depict the standard curves use to estimate the amount of each mRNA present in the two groups. The curves were generated using serial dilutions of reverse transcription reactions derived from one μg of total RNA from 7-day-old mouse ovaries (equivalent to 10 −1 – 10 −8 ) using an ABI Prism 7700 sequence Detection System (Perkin Elmer Applied Biosystems), in a total volume of 10 μl, containing 2 μl of reverse transcription reaction, 5 μl of TaqMan Universal PCR Master Mix (Perkin Elmer Applied Biosystems), and 3 μl primer mix (250 nM of each gene-specific and ribosomal fluorescent probes and 300 nM of each gene-specific unlabeled primer plus 80 nM of each unlabeled ribosomal primer). (C and D) Ovarian content of GDF-9 (C) and KL (D) mRNA detected by RealTime PCR in wild-type and trkB −/− 7-day-old mice. (E and F) Representative ethidium bromide-stained gels showing BMPRII (E) and c-kit (F) PCR products amplified from total ovarian RNA using the Relative Quantitative PCR conditions described in Materials and methods. (G and H) Relative content of BMPRII (G) and c-kit (H) mRNAs estimated by Relative Quantitative PCR. Values are shown as ratios between the gene-specific signal and the signal provided by 18S ribosomal RNA coamplified with each gene of interest. Bars are mean ± SEM and numbers on top of bars are number of mice per group. Because the KL and BMPRII mRNAs are expressed in granulosa cells, the values obtained in trkB −/− mice were corrected to take into consideration the reduction in granulosa cell number observed in these animals.

    Journal: Developmental biology

    Article Title: TrkB receptors are required for follicular growth and oocyte survival in the mammalian ovary

    doi: 10.1016/j.ydbio.2003.12.001

    Figure Lengend Snippet: The ovarian content of the mRNAs encoding GDF-9 and its presumed receptor, BMPRII, and KL and its c-kit receptor is not altered in trkB −/− mice. (A and B) Representative RealTime PCR amplification profiles of GDF-9 mRNA (A) and KL mRNA (B) from the ovary of trkB −/− mice compared with that of wild-type littermates. The insets depict the standard curves use to estimate the amount of each mRNA present in the two groups. The curves were generated using serial dilutions of reverse transcription reactions derived from one μg of total RNA from 7-day-old mouse ovaries (equivalent to 10 −1 – 10 −8 ) using an ABI Prism 7700 sequence Detection System (Perkin Elmer Applied Biosystems), in a total volume of 10 μl, containing 2 μl of reverse transcription reaction, 5 μl of TaqMan Universal PCR Master Mix (Perkin Elmer Applied Biosystems), and 3 μl primer mix (250 nM of each gene-specific and ribosomal fluorescent probes and 300 nM of each gene-specific unlabeled primer plus 80 nM of each unlabeled ribosomal primer). (C and D) Ovarian content of GDF-9 (C) and KL (D) mRNA detected by RealTime PCR in wild-type and trkB −/− 7-day-old mice. (E and F) Representative ethidium bromide-stained gels showing BMPRII (E) and c-kit (F) PCR products amplified from total ovarian RNA using the Relative Quantitative PCR conditions described in Materials and methods. (G and H) Relative content of BMPRII (G) and c-kit (H) mRNAs estimated by Relative Quantitative PCR. Values are shown as ratios between the gene-specific signal and the signal provided by 18S ribosomal RNA coamplified with each gene of interest. Bars are mean ± SEM and numbers on top of bars are number of mice per group. Because the KL and BMPRII mRNAs are expressed in granulosa cells, the values obtained in trkB −/− mice were corrected to take into consideration the reduction in granulosa cell number observed in these animals.

    Article Snippet: Equal volumes of the PCR reactions were electrophoresed on 2% agarose gels stained with ethidium bromide, the gels were imaged in a Gel Doc 2000 (Bio-Rad Laboratories, Hercules, CA), and the images were quantitated using the image analysis Quantity One software (Bio-Rad Laboratories).

    Techniques: Mouse Assay, Polymerase Chain Reaction, Amplification, Generated, Derivative Assay, Sequencing, Staining, Real-time Polymerase Chain Reaction

    Study of flavone–DNA interaction. ( A ) A change in the emission spectra (λ excitation = 364 nm) of 50 µM baicalein was determined as a function of CT DNA concentration in the range of 0–0.22 mM. Arrow indicates the change in the emission spectra of baicalein on addition of CT DNA. Control fluorescence spectrum of CT DNA is indicated. ( B ) Flavones–DNA interaction as studied by agarose gel electrophoresis. Lane 2, relaxed pBS (SK + ) DNA generated by treatment of pBS (SK + ) DNA (lane 1) with excess of topoisomerase I, followed by phenol–chloroform extraction and ethanol precipitation. Lanes 3–6, same as lane 2, but incubated with 50 and 300 µM of m -AMSA and etoposide, respectively, as indicated. Lanes 7–15, same as lane 2, but incubated with 50, 100, 300 µM of baicalein, luteolin and quercetin as indicated. ( C ) Fluorescence-based ethidium bromide displacement assay. Samples contained 1 µM of EtBr and 5 nM CT DNA. Increasing concentration (0–300 µM) of baicalein, luteolin, quercetin, m -AMSA and etoposide were added as indicated. EtBr fluorescence was monitored with excitation wavelength at 510 nm and emission at 590 nm.

    Journal: Nucleic Acids Research

    Article Title: Differential induction of Leishmania donovani bi-subunit topoisomerase I-DNA cleavage complex by selected flavones and camptothecin: activity of flavones against camptothecin-resistant topoisomerase I

    doi: 10.1093/nar/gkj502

    Figure Lengend Snippet: Study of flavone–DNA interaction. ( A ) A change in the emission spectra (λ excitation = 364 nm) of 50 µM baicalein was determined as a function of CT DNA concentration in the range of 0–0.22 mM. Arrow indicates the change in the emission spectra of baicalein on addition of CT DNA. Control fluorescence spectrum of CT DNA is indicated. ( B ) Flavones–DNA interaction as studied by agarose gel electrophoresis. Lane 2, relaxed pBS (SK + ) DNA generated by treatment of pBS (SK + ) DNA (lane 1) with excess of topoisomerase I, followed by phenol–chloroform extraction and ethanol precipitation. Lanes 3–6, same as lane 2, but incubated with 50 and 300 µM of m -AMSA and etoposide, respectively, as indicated. Lanes 7–15, same as lane 2, but incubated with 50, 100, 300 µM of baicalein, luteolin and quercetin as indicated. ( C ) Fluorescence-based ethidium bromide displacement assay. Samples contained 1 µM of EtBr and 5 nM CT DNA. Increasing concentration (0–300 µM) of baicalein, luteolin, quercetin, m -AMSA and etoposide were added as indicated. EtBr fluorescence was monitored with excitation wavelength at 510 nm and emission at 590 nm.

    Article Snippet: The amount of supercoiled monomer DNA band florescence after ethidium bromide (EtBr) (0.5 µg/ml) staining was quantitated by using Gel Doc 2000 under UV illumination (Bio Rad-Quality one software).

    Techniques: Concentration Assay, Fluorescence, Agarose Gel Electrophoresis, Generated, Ethanol Precipitation, Incubation

    The effect of temperature on tHDA enzymatic activity . (A) The influence of temperature on the enzymatic activity of the isothermal tHDA system. Samples were incubated at 65°C (lane 2), 37°C (lane 3), 23°C (lane 4), and 4°C (lane 5) for 1.5 h. Lane 1 contains a 50 bp DNA ladder. The 1350 bp, 100 bp, and 50 bp bands are labeled. (B) The influence of overnight incubation temperature on the enzymatic activity of the isothermal tHDA system. Samples were incubated at 4°C (lane 2) and 23°C (lane 3) overnight prior to incubation at 65°C for 1.5 h. Lane 1 is a 50 bp DNA ladder. Reaction products were observed by ethidium bromide staining of a 1.8% agarose gel.

    Journal: BMC Research Notes

    Article Title: Intravesicle Isothermal DNA Replication

    doi: 10.1186/1756-0500-4-128

    Figure Lengend Snippet: The effect of temperature on tHDA enzymatic activity . (A) The influence of temperature on the enzymatic activity of the isothermal tHDA system. Samples were incubated at 65°C (lane 2), 37°C (lane 3), 23°C (lane 4), and 4°C (lane 5) for 1.5 h. Lane 1 contains a 50 bp DNA ladder. The 1350 bp, 100 bp, and 50 bp bands are labeled. (B) The influence of overnight incubation temperature on the enzymatic activity of the isothermal tHDA system. Samples were incubated at 4°C (lane 2) and 23°C (lane 3) overnight prior to incubation at 65°C for 1.5 h. Lane 1 is a 50 bp DNA ladder. Reaction products were observed by ethidium bromide staining of a 1.8% agarose gel.

    Article Snippet: Ethidium bromide gels were visualized with either a BioDoc-It Imaging System (UVP) or a Molecular Imager ChemiDoc XRS System (Bio-Rad).

    Techniques: Activity Assay, Incubation, Labeling, Staining, Agarose Gel Electrophoresis

    The influence of freeze/thaw cycles on tHDA enzymatic activity . Unencapsulated reaction mixtures were subjected to either 0 (lane 2), 5 (lane 3), 10 (lane 4), or 20 (lane 5) cycles of freeze-thawing. Lane 1 is a 50 bp DNA ladder. The 1350 bp, 100 bp, and 50 bp bands are labeled. Reaction products were visualized by ethidium bromide staining of a 1.8% agarose gel. The full length reaction product is 85 bp.

    Journal: BMC Research Notes

    Article Title: Intravesicle Isothermal DNA Replication

    doi: 10.1186/1756-0500-4-128

    Figure Lengend Snippet: The influence of freeze/thaw cycles on tHDA enzymatic activity . Unencapsulated reaction mixtures were subjected to either 0 (lane 2), 5 (lane 3), 10 (lane 4), or 20 (lane 5) cycles of freeze-thawing. Lane 1 is a 50 bp DNA ladder. The 1350 bp, 100 bp, and 50 bp bands are labeled. Reaction products were visualized by ethidium bromide staining of a 1.8% agarose gel. The full length reaction product is 85 bp.

    Article Snippet: Ethidium bromide gels were visualized with either a BioDoc-It Imaging System (UVP) or a Molecular Imager ChemiDoc XRS System (Bio-Rad).

    Techniques: Activity Assay, Labeling, Staining, Agarose Gel Electrophoresis

    Chitosan nanoparticle temporal stability. Stability was assessed at 0.5, 4, and 24 hours after complex formation using polyacrylamide gel electrophoresis at a pH of 6.5 (MES 1X) and pH8 (TAE 1X). Chitosan 92-10 at different N:P ratios (0.5, 2, and 10) was complexed with: ( A ) dsODN-RecQL1 at a pH of 6.5; ( B ) dsODN-RecQL1 at a pH of 8; ( C ) dsODN-ApoB at a pH of 6.5, and ( D ) dsODN-ApoB at a pH of 8. Unstable nanoparticles release dsODNs which become visible following EtBr staining on polyacrylamide gel following ethidium bromide staining of the polyacrylamide gel. Abbreviations: ApoB, apolipoprotein B; dsODN, double-stranded oligodeoxynucleotide; N:P, amine to phosphate.

    Journal: International Journal of Nanomedicine

    Article Title: Low molecular weight chitosan nanoparticulate system at low N:P ratio for nontoxic polynucleotide delivery

    doi: 10.2147/IJN.S26571

    Figure Lengend Snippet: Chitosan nanoparticle temporal stability. Stability was assessed at 0.5, 4, and 24 hours after complex formation using polyacrylamide gel electrophoresis at a pH of 6.5 (MES 1X) and pH8 (TAE 1X). Chitosan 92-10 at different N:P ratios (0.5, 2, and 10) was complexed with: ( A ) dsODN-RecQL1 at a pH of 6.5; ( B ) dsODN-RecQL1 at a pH of 8; ( C ) dsODN-ApoB at a pH of 6.5, and ( D ) dsODN-ApoB at a pH of 8. Unstable nanoparticles release dsODNs which become visible following EtBr staining on polyacrylamide gel following ethidium bromide staining of the polyacrylamide gel. Abbreviations: ApoB, apolipoprotein B; dsODN, double-stranded oligodeoxynucleotide; N:P, amine to phosphate.

    Article Snippet: Samples were migrated at 90 V during 1 hour then stained with 0.5 μg/mL ethidium bromide solution before visualization.

    Techniques: Polyacrylamide Gel Electrophoresis, Staining

    Representative PCR picture showing amplification of TLR9 gene segment for C296T/ Pro99Leu gene polymorphism on ethidium bromide-stained 2% agarose gel. Lane M is 100 bp molecular marker (Takara, Japan; Cat# RR820A), Lane 1 is negative control and Lanes 2–7 are tumor DNA showing PCR products of 337 bp. (Abbreviations: PCR, Polymerase Chain Reaction; TLR9, Toll-like receptor 9; bp, base pair).

    Journal: F1000Research

    Article Title: Absence of toll-like receptor 9 Pro99Leu polymorphism in cervical cancer

    doi: 10.12688/f1000research.14840.2

    Figure Lengend Snippet: Representative PCR picture showing amplification of TLR9 gene segment for C296T/ Pro99Leu gene polymorphism on ethidium bromide-stained 2% agarose gel. Lane M is 100 bp molecular marker (Takara, Japan; Cat# RR820A), Lane 1 is negative control and Lanes 2–7 are tumor DNA showing PCR products of 337 bp. (Abbreviations: PCR, Polymerase Chain Reaction; TLR9, Toll-like receptor 9; bp, base pair).

    Article Snippet: The quality and quantity of DNA was determined using ethidium bromide-stained 1% agarose gel on GelDoc system (BioRad, USA) as well as a NanoDrop 2000 (Thermofisher, USA).

    Techniques: Polymerase Chain Reaction, Amplification, Staining, Agarose Gel Electrophoresis, Marker, Negative Control

    The transcripts for the enterotoxin genes act and alt were eliminated in the indicated mutants, based on Northern blot analysis. Total RNA from wild-type A. hydrophila and its enterotoxin gene-deficient mutants was isolated and hybridized with the act gene probe (A-I) and the alt gene probe (B-I), as described in Materials and Methods. Lane 1, wild-type A. hydrophila ; lane 2, mutant SSUΔ act ; lane 3, mutant SSUΔ alt ; lane 4, mutant SSUΔ ast ; lane 5, mutant SSUΔ alt , ast ; lane 6, mutant SSUΔ act , alt ; lane 7, mutant SSUΔ act , ast ; lane 8, mutant SSUΔ act , alt , ast . The RNA loaded in each lane was quantitated by scanning 16S and 23S rRNA bands after ethidium bromide staining of the gel (A-II and B-II).

    Journal: Infection and Immunity

    Article Title: Role of Various Enterotoxins in Aeromonas hydrophila-Induced Gastroenteritis: Generation of Enterotoxin Gene-Deficient Mutants and Evaluation of Their Enterotoxic Activity

    doi: 10.1128/IAI.70.4.1924-1935.2002

    Figure Lengend Snippet: The transcripts for the enterotoxin genes act and alt were eliminated in the indicated mutants, based on Northern blot analysis. Total RNA from wild-type A. hydrophila and its enterotoxin gene-deficient mutants was isolated and hybridized with the act gene probe (A-I) and the alt gene probe (B-I), as described in Materials and Methods. Lane 1, wild-type A. hydrophila ; lane 2, mutant SSUΔ act ; lane 3, mutant SSUΔ alt ; lane 4, mutant SSUΔ ast ; lane 5, mutant SSUΔ alt , ast ; lane 6, mutant SSUΔ act , alt ; lane 7, mutant SSUΔ act , ast ; lane 8, mutant SSUΔ act , alt , ast . The RNA loaded in each lane was quantitated by scanning 16S and 23S rRNA bands after ethidium bromide staining of the gel (A-II and B-II).

    Article Snippet: The amount of RNA in each lane was quantitated by scanning 23S or 16S rRNA bands after ethidium bromide staining of the gel, using the Gel Doc 2000 System (Bio-Rad Laboratories, Hercules, Calif.).

    Techniques: Activated Clotting Time Assay, Northern Blot, Isolation, Mutagenesis, AST Assay, Staining

    LmrP mediates cellular sensitivity to Hoechst 33342 but resistance to ethidium. Growth rate of LmrP-expressing lactococcal cells and cells without LmrP expression (control) in the presence of up to 8.4 μM Hoechst 33342 (A), or up to 24 μM ethidium (B) was determined relative to the maximum growth rate in the absence of drug. The error bars for some of the data points were too small to be displayed, and are hidden behind the data point symbols.

    Journal: PLoS ONE

    Article Title: Hoechst 33342 Is a Hidden “Janus” amongst Substrates for the Multidrug Efflux Pump LmrP

    doi: 10.1371/journal.pone.0141991

    Figure Lengend Snippet: LmrP mediates cellular sensitivity to Hoechst 33342 but resistance to ethidium. Growth rate of LmrP-expressing lactococcal cells and cells without LmrP expression (control) in the presence of up to 8.4 μM Hoechst 33342 (A), or up to 24 μM ethidium (B) was determined relative to the maximum growth rate in the absence of drug. The error bars for some of the data points were too small to be displayed, and are hidden behind the data point symbols.

    Article Snippet: Glucose (25 mM) was added 3 min before the addition of 0.1 μM Hoechst 33342 (trihydrochloride salt, fluoropure grade, Molecular Probes, Thermofisher Scientific) or 2 μM ethidium (bromide salt, molecular grade, Promega).

    Techniques: Expressing

    LmrP mediates active efflux of ethidium. The efflux experiments described in Fig 3 were performed with ethidium at a final concentration of 2 μM. Traces represent data obtained in three independent experiments using different batches of cells.

    Journal: PLoS ONE

    Article Title: Hoechst 33342 Is a Hidden “Janus” amongst Substrates for the Multidrug Efflux Pump LmrP

    doi: 10.1371/journal.pone.0141991

    Figure Lengend Snippet: LmrP mediates active efflux of ethidium. The efflux experiments described in Fig 3 were performed with ethidium at a final concentration of 2 μM. Traces represent data obtained in three independent experiments using different batches of cells.

    Article Snippet: Glucose (25 mM) was added 3 min before the addition of 0.1 μM Hoechst 33342 (trihydrochloride salt, fluoropure grade, Molecular Probes, Thermofisher Scientific) or 2 μM ethidium (bromide salt, molecular grade, Promega).

    Techniques: Concentration Assay

    LmrP expressing cells show reduced accumulation of ethidium compared to control cells. The substrate accumulation experiments described in Fig 1A were performed with ethidium at a final concentration of 2 μM. Traces represent data obtained in three independent experiments using different batches of cells.

    Journal: PLoS ONE

    Article Title: Hoechst 33342 Is a Hidden “Janus” amongst Substrates for the Multidrug Efflux Pump LmrP

    doi: 10.1371/journal.pone.0141991

    Figure Lengend Snippet: LmrP expressing cells show reduced accumulation of ethidium compared to control cells. The substrate accumulation experiments described in Fig 1A were performed with ethidium at a final concentration of 2 μM. Traces represent data obtained in three independent experiments using different batches of cells.

    Article Snippet: Glucose (25 mM) was added 3 min before the addition of 0.1 μM Hoechst 33342 (trihydrochloride salt, fluoropure grade, Molecular Probes, Thermofisher Scientific) or 2 μM ethidium (bromide salt, molecular grade, Promega).

    Techniques: Expressing, Concentration Assay

    NAI or NAI-imine increase P2X7-mediated IL-1 β release but not ethidium + uptake. LPS-primed J774 cells in physiological medium were preincubated in the presence of DMSO, or 1 μ M NAI or 1 μ M NAI-imine for 30 min, and then in the absence or presence of 3 mM ATP for ((a) and (b)) 20 min or (c) 5 min in the presence of 25 μ M ethidium bromide. (a) The IL-1 β concentration in cell-free supernatants was measured using an ELISA. (b) The LDH activity in cell-free supernatants and cell lysates was measured using a cytotoxicity kit, and results are expressed as a percentage of maximal LDH release from lysed cells. (c) Ethidium + uptake was measured by flow cytometry and is expressed as mean fluorescence intensity (MFI). ((a) to (c)) Results are mean ± SD ( n = 3); ∗ P

    Journal: Mediators of Inflammation

    Article Title: N-Alkyl-Substituted Isatins Enhance P2X7 Receptor-Induced Interleukin-1β Release from Murine Macrophages

    doi: 10.1155/2016/2097219

    Figure Lengend Snippet: NAI or NAI-imine increase P2X7-mediated IL-1 β release but not ethidium + uptake. LPS-primed J774 cells in physiological medium were preincubated in the presence of DMSO, or 1 μ M NAI or 1 μ M NAI-imine for 30 min, and then in the absence or presence of 3 mM ATP for ((a) and (b)) 20 min or (c) 5 min in the presence of 25 μ M ethidium bromide. (a) The IL-1 β concentration in cell-free supernatants was measured using an ELISA. (b) The LDH activity in cell-free supernatants and cell lysates was measured using a cytotoxicity kit, and results are expressed as a percentage of maximal LDH release from lysed cells. (c) Ethidium + uptake was measured by flow cytometry and is expressed as mean fluorescence intensity (MFI). ((a) to (c)) Results are mean ± SD ( n = 3); ∗ P

    Article Snippet: Dimethyl sulfoxide (DMSO) and ethidium bromide were from Amresco (Solon, USA).

    Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Flow Cytometry, Cytometry, Fluorescence

    Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) ethidium bromide; ( C ) YO; ( D ) YOYO.

    Journal: Biophysical Journal

    Article Title: Identification of Binding Mechanisms in Single Molecule-DNA Complexes

    doi:

    Figure Lengend Snippet: Force-extension traces for the complexes of poly(dG-dC) dsDNA with the intercalants ( A ) daunomycin; ( B ) ethidium bromide; ( C ) YO; ( D ) YOYO.

    Article Snippet: The DNA-binding agents daunomycin (Sigma), ethidium bromide (Merck, Darmstadt, Germany), distamycin A (Sigma), YO (Molecular Probes, Eugene, OR), YOYO (Molecular Probes), Ac-(Leu-Ala-Arg-Leu)3 -NH-linker and Ac-(Aib-Leu-Arg)4 -NH-linker were added to 10 μ l of the DNA solution in a concentration of 150 μ M, corresponding to a 1:10 ratio of agent molecules per base pair.

    Techniques: