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  • 99
    Thermo Fisher egta
    Filamin A is locally translated in injured axons. A , C , E , and G , Western blots of control (− Ax ) and axotomized (+ Ax ) sciatic nerves treated with vehicle or cycloheximide ( CHX , A ), <t>EGTA</t> ( C ), <t>Gö6976</t> ( E ), and H-89 ( G ). B , D , F , and H , quantifications of filamin A levels in A , C , E , and G . Data are mean ± S.E. n = 3; ***, p
    Egta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3780 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore • egta
    Filamin A is locally translated in injured axons. A , C , E , and G , Western blots of control (− Ax ) and axotomized (+ Ax ) sciatic nerves treated with vehicle or cycloheximide ( CHX , A ), <t>EGTA</t> ( C ), <t>Gö6976</t> ( E ), and H-89 ( G ). B , D , F , and H , quantifications of filamin A levels in A , C , E , and G . Data are mean ± S.E. n = 3; ***, p
    • Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    FUJIFILM egtazic acid
    Filamin A is locally translated in injured axons. A , C , E , and G , Western blots of control (− Ax ) and axotomized (+ Ax ) sciatic nerves treated with vehicle or cycloheximide ( CHX , A ), <t>EGTA</t> ( C ), <t>Gö6976</t> ( E ), and H-89 ( G ). B , D , F , and H , quantifications of filamin A levels in A , C , E , and G . Data are mean ± S.E. n = 3; ***, p
    Egtazic Acid, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore egta
    <t>PABA/NO</t> treatment of HL60 cells results in temporal and dose-dependent increases in intracellular Ca 2+ . A) Time-dependent increases were assessed following treatment with 20 nM ThG; 25 µM PABA/NO; 25 µM PABA; 20 nM ThG after pretreatment with PABA/NO (15 µM, 30 min); 25 µM DEA NONOate B) Competitive inhibition of ThG (100 nM) - mediated increases in intracellular Ca 2+ by PABA/NO, purified homogeneous nitro-aromatic product (MW 622 Da), or PABA. C) Dose-dependent increases in Ca 2+ following PABA/NO (Control) or PABA were measured; in the presence of the intracellular Ca 2+ chelator BAPTA-AM (5 µM); or in the presence of the extracellular Ca 2+ chelator, <t>EGTA</t> (5 mM, long dashedline). The fluorescence measurements were recalculated as actual intracellular Ca 2+ concentrations (see Materials and Methods ). D) Kinetics of intracellular NO (left Y-axis) and Ca 2+ (right Y-axis, solid line) generation after PABA/NO (15 µM) addition. Data are the average representative traces (A and D) or mean ± SE (B, C) for 3 independent experiments.
    Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Boston BioProducts egta
    <t>PABA/NO</t> treatment of HL60 cells results in temporal and dose-dependent increases in intracellular Ca 2+ . A) Time-dependent increases were assessed following treatment with 20 nM ThG; 25 µM PABA/NO; 25 µM PABA; 20 nM ThG after pretreatment with PABA/NO (15 µM, 30 min); 25 µM DEA NONOate B) Competitive inhibition of ThG (100 nM) - mediated increases in intracellular Ca 2+ by PABA/NO, purified homogeneous nitro-aromatic product (MW 622 Da), or PABA. C) Dose-dependent increases in Ca 2+ following PABA/NO (Control) or PABA were measured; in the presence of the intracellular Ca 2+ chelator BAPTA-AM (5 µM); or in the presence of the extracellular Ca 2+ chelator, <t>EGTA</t> (5 mM, long dashedline). The fluorescence measurements were recalculated as actual intracellular Ca 2+ concentrations (see Materials and Methods ). D) Kinetics of intracellular NO (left Y-axis) and Ca 2+ (right Y-axis, solid line) generation after PABA/NO (15 µM) addition. Data are the average representative traces (A and D) or mean ± SE (B, C) for 3 independent experiments.
    Egta, supplied by Boston BioProducts, used in various techniques. Bioz Stars score: 95/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Carl Roth GmbH egta
    Effect of PF-IgGs on binding of Dsg1-coated beads to <t>HaCaT</t> cells. Beads were allowed to settle on the surface of HaCaT cells for 30 minutes (control). Number of bound beads was reduced by simultaneous incubation of <t>EGTA</t> (5 mM, 30 minutes). Incubation of monolayers with attached beads for an additional 30 minutes with IgG fractions from 2 patients (PF1- and PF2-IgG, 35 μg/ml each) as well as with a monoclonal antibody (1:50) directed against the extracellular domain of human Dsg1 significantly reduced the number of bound beads (label in white box and white bars). Immunoabsorption using Dsg1-coated beads but not control absorption using VE-cadherin–labeled beads completely abolished the effect of PF-IgGs on bead adhesion. Preincubation of HaCaT cells with PF-IgGs prior to bead settlement also reduced bead binding (label in gray box and gray bars) whereas preincubation of beads with PF-IgGs did not inhibit bead binding. In contrast, the monoclonal Dsg1 antibody reduced bead binding also when applied for preincubation with beads, indicating a different mechanism underlying the reduction of Dsg1 adhesion ( n = 6 for each condition).
    Egta, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 96/100, based on 107 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Dojindo Labs egta
    ATP's antimicrobial effect is attributable to its iron-chelating activity. (a) Blocking effects of MgCl 2 and FeCl 3 on ATP's anti- M. intracellulare antimicrobial activity. (b) Anti- M. intracellulare activities of ATP and various metal-chelating agents (pyrophosphate (PPi), <t>EDTA,</t> <t>EGTA)</t> and blocking effects of MgCl 2 . (c) Anti- S. aureus activities of ATP and various metal-chelating agents. (d) Ferric ion-chelating activities of ATP and EDTA measured by CAS assay. (e) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against M. intracellulare . (f) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against P. aeruginosa . (g) Siderophore production by ATP-resistant E. coli during the course of cultivation. (h) Siderophore production by two ATP-resistant K. pneumoniae strains during 48-h cultivation in the presence of ATP. (i) Siderophore production by ATP-susceptible S. aureus and ATP-resistant E. coli during 24-h cultivation in the presence of ATP (5 mM). (j) Siderophore production by ATP-susceptible and ATP-resistant strains of P. aeruginosa during 24-h cultivation.
    Egta, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 94/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Fisher Scientific egta
    Typical force versus relative deformation profiles for <t>MDA-MB-468</t> cells labeled with various dyes: a control cell (1, red), a <t>EGTA</t> treated cell (2, gray), and cells labeled with 5 μM CFDA-SE (3, blue), CMFDA (4, green), CMTMR (5, orange) and Calcein
    Egta, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 95/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Fluka Chemical egta
    Typical force versus relative deformation profiles for <t>MDA-MB-468</t> cells labeled with various dyes: a control cell (1, red), a <t>EGTA</t> treated cell (2, gray), and cells labeled with 5 μM CFDA-SE (3, blue), CMFDA (4, green), CMTMR (5, orange) and Calcein
    Egta, supplied by Fluka Chemical, used in various techniques. Bioz Stars score: 81/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Amresco egta
    Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L <t>BAPTA-AM</t> + 0.5 mmol/L <t>EGTA,</t> n = 5) or introducing a
    Egta, supplied by Amresco, used in various techniques. Bioz Stars score: 93/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Euromedex egta
    Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L <t>BAPTA-AM</t> + 0.5 mmol/L <t>EGTA,</t> n = 5) or introducing a
    Egta, supplied by Euromedex, used in various techniques. Bioz Stars score: 95/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    RPI Inc egta
    Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L <t>BAPTA-AM</t> + 0.5 mmol/L <t>EGTA,</t> n = 5) or introducing a
    Egta, supplied by RPI Inc, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Valiant egta
    Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L <t>BAPTA-AM</t> + 0.5 mmol/L <t>EGTA,</t> n = 5) or introducing a
    Egta, supplied by Valiant, used in various techniques. Bioz Stars score: 93/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egta  (Tocris)
    99
    Tocris egta
    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM <t>EGTA</t> or 10 μM <t>BAPTA-AM,</t> for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P
    Egta, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 199 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Applichem egta
    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM <t>EGTA</t> or 10 μM <t>BAPTA-AM,</t> for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P
    Egta, supplied by Applichem, used in various techniques. Bioz Stars score: 89/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Beijing Solarbio Science egta
    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM <t>EGTA</t> or 10 μM <t>BAPTA-AM,</t> for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P
    Egta, supplied by Beijing Solarbio Science, 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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    93
    Boehringer Mannheim egta
    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM <t>EGTA</t> or 10 μM <t>BAPTA-AM,</t> for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P
    Egta, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 93/100, based on 778 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Nacalai egta
    Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and <t>EGTA</t> or <t>EDTA</t> was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the
    Egta, supplied by Nacalai, used in various techniques. Bioz Stars score: 97/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Promega egta
    Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and <t>EGTA</t> or <t>EDTA</t> was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the
    Egta, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 276 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BioShop egta
    Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and <t>EGTA</t> or <t>EDTA</t> was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the
    Egta, supplied by BioShop, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Fisher Bioreagents egta
    Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and <t>EGTA</t> or <t>EDTA</t> was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the
    Egta, supplied by Fisher Bioreagents, used in various techniques. Bioz Stars score: 88/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck & Co egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Merck & Co, used in various techniques. Bioz Stars score: 93/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Merck KGaA egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 96/100, based on 129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egta  (TaKaRa)
    97
    TaKaRa egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by TaKaRa, used in various techniques. Bioz Stars score: 97/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 830 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egta  (Lonza)
    88
    Lonza egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Lonza, used in various techniques. Bioz Stars score: 88/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Santa Cruz Biotechnology egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 304 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Trion Research egta
    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM <t>EGTA,</t> 5 mM Ni 2+ , 125 μM verapamil (VPL), or control <t>HBSS.</t> Results are means ± SEM for four to six experiments. ∗, P
    Egta, supplied by Trion Research, used in various techniques. Bioz Stars score: 91/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    AAT Bioquest egta
    Excessive Ca 2+ -Independent Actomyosin Associations during Diastole Promote Enhanced Myocyte Shortening and Incomplete Relaxation of Hand > Act57B A295S Cardiomyocytes (A) M modes generated from the same region of a three-week-old Hand > Act57B WT heart reveal graded responses of cardiac diameters following exposure to distinct small-molecule compounds. Red arrowheads indicate the position of the heart wall edges during diastole, upon incubation with <t>EGTA/EGTA,AM,</t> and finally, following the addition of blebbistatin. Incubation with EGTA/EGTA,AM resulted in complete cessation of wall motion. Relative to the diameter during diastole, each treatment induced a slight increase in diameter across the heart tube. (B) Significant, incremental increases in cardiac diameters were verified for all genotypes following extra- and intracellular Ca 2+ chelation and upon blebbistatin incubation. (C) The average change in diameter across the heart wall in response to EGTA/EGTA,AM was similar among all lines. (D) Blebbistatin treatment prompted a significantly greater response across the wall of Hand > Act57B A295S hearts relative to that observed for Hand × yw and Hand > Act57B WT hearts. Data are presented as mean ± SEM (n = 21). ***p ≤ 0.001. See also Figure S3 .
    Egta, supplied by AAT Bioquest, used in various techniques. Bioz Stars score: 96/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Fluka Chemie egta
    Excessive Ca 2+ -Independent Actomyosin Associations during Diastole Promote Enhanced Myocyte Shortening and Incomplete Relaxation of Hand > Act57B A295S Cardiomyocytes (A) M modes generated from the same region of a three-week-old Hand > Act57B WT heart reveal graded responses of cardiac diameters following exposure to distinct small-molecule compounds. Red arrowheads indicate the position of the heart wall edges during diastole, upon incubation with <t>EGTA/EGTA,AM,</t> and finally, following the addition of blebbistatin. Incubation with EGTA/EGTA,AM resulted in complete cessation of wall motion. Relative to the diameter during diastole, each treatment induced a slight increase in diameter across the heart tube. (B) Significant, incremental increases in cardiac diameters were verified for all genotypes following extra- and intracellular Ca 2+ chelation and upon blebbistatin incubation. (C) The average change in diameter across the heart wall in response to EGTA/EGTA,AM was similar among all lines. (D) Blebbistatin treatment prompted a significantly greater response across the wall of Hand > Act57B A295S hearts relative to that observed for Hand × yw and Hand > Act57B WT hearts. Data are presented as mean ± SEM (n = 21). ***p ≤ 0.001. See also Figure S3 .
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    Filamin A is locally translated in injured axons. A , C , E , and G , Western blots of control (− Ax ) and axotomized (+ Ax ) sciatic nerves treated with vehicle or cycloheximide ( CHX , A ), EGTA ( C ), Gö6976 ( E ), and H-89 ( G ). B , D , F , and H , quantifications of filamin A levels in A , C , E , and G . Data are mean ± S.E. n = 3; ***, p

    Journal: The Journal of Biological Chemistry

    Article Title: Filamin A Is Required in Injured Axons for HDAC5 Activity and Axon Regeneration *

    doi: 10.1074/jbc.M115.638445

    Figure Lengend Snippet: Filamin A is locally translated in injured axons. A , C , E , and G , Western blots of control (− Ax ) and axotomized (+ Ax ) sciatic nerves treated with vehicle or cycloheximide ( CHX , A ), EGTA ( C ), Gö6976 ( E ), and H-89 ( G ). B , D , F , and H , quantifications of filamin A levels in A , C , E , and G . Data are mean ± S.E. n = 3; ***, p

    Article Snippet: For sciatic nerve delivery of chemical compounds, the chemicals cycloheximide (1 mg/kg, Sigma), EGTA (10 m m , Life Technologies), Gö6976 (1 mg/kg, Tocris), and H-89 (1 mg/kg, Tocris) were dissolved in dimethyl sulfoxide.

    Techniques: Western Blot

    PABA/NO treatment of HL60 cells results in temporal and dose-dependent increases in intracellular Ca 2+ . A) Time-dependent increases were assessed following treatment with 20 nM ThG; 25 µM PABA/NO; 25 µM PABA; 20 nM ThG after pretreatment with PABA/NO (15 µM, 30 min); 25 µM DEA NONOate B) Competitive inhibition of ThG (100 nM) - mediated increases in intracellular Ca 2+ by PABA/NO, purified homogeneous nitro-aromatic product (MW 622 Da), or PABA. C) Dose-dependent increases in Ca 2+ following PABA/NO (Control) or PABA were measured; in the presence of the intracellular Ca 2+ chelator BAPTA-AM (5 µM); or in the presence of the extracellular Ca 2+ chelator, EGTA (5 mM, long dashedline). The fluorescence measurements were recalculated as actual intracellular Ca 2+ concentrations (see Materials and Methods ). D) Kinetics of intracellular NO (left Y-axis) and Ca 2+ (right Y-axis, solid line) generation after PABA/NO (15 µM) addition. Data are the average representative traces (A and D) or mean ± SE (B, C) for 3 independent experiments.

    Journal: PLoS ONE

    Article Title: Diazeniumdiolate Mediated Nitrosative Stress Alters Nitric Oxide Homeostasis through Intracellular Calcium and S-Glutathionylation of Nitric Oxide Synthetase

    doi: 10.1371/journal.pone.0014151

    Figure Lengend Snippet: PABA/NO treatment of HL60 cells results in temporal and dose-dependent increases in intracellular Ca 2+ . A) Time-dependent increases were assessed following treatment with 20 nM ThG; 25 µM PABA/NO; 25 µM PABA; 20 nM ThG after pretreatment with PABA/NO (15 µM, 30 min); 25 µM DEA NONOate B) Competitive inhibition of ThG (100 nM) - mediated increases in intracellular Ca 2+ by PABA/NO, purified homogeneous nitro-aromatic product (MW 622 Da), or PABA. C) Dose-dependent increases in Ca 2+ following PABA/NO (Control) or PABA were measured; in the presence of the intracellular Ca 2+ chelator BAPTA-AM (5 µM); or in the presence of the extracellular Ca 2+ chelator, EGTA (5 mM, long dashedline). The fluorescence measurements were recalculated as actual intracellular Ca 2+ concentrations (see Materials and Methods ). D) Kinetics of intracellular NO (left Y-axis) and Ca 2+ (right Y-axis, solid line) generation after PABA/NO (15 µM) addition. Data are the average representative traces (A and D) or mean ± SE (B, C) for 3 independent experiments.

    Article Snippet: PABA (4-aminobenzoic acid sodium salt), TCEP (tris(2-carboxyethyl)phosphine), W-7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide), 2,2′-(Hydroxynitrosohydrazono)bis-ethanimine (DETA/NO), and EGTA were purchased from Sigma (St. Louis, MO).

    Techniques: Inhibition, Purification, Fluorescence

    Effect of PF-IgGs on binding of Dsg1-coated beads to HaCaT cells. Beads were allowed to settle on the surface of HaCaT cells for 30 minutes (control). Number of bound beads was reduced by simultaneous incubation of EGTA (5 mM, 30 minutes). Incubation of monolayers with attached beads for an additional 30 minutes with IgG fractions from 2 patients (PF1- and PF2-IgG, 35 μg/ml each) as well as with a monoclonal antibody (1:50) directed against the extracellular domain of human Dsg1 significantly reduced the number of bound beads (label in white box and white bars). Immunoabsorption using Dsg1-coated beads but not control absorption using VE-cadherin–labeled beads completely abolished the effect of PF-IgGs on bead adhesion. Preincubation of HaCaT cells with PF-IgGs prior to bead settlement also reduced bead binding (label in gray box and gray bars) whereas preincubation of beads with PF-IgGs did not inhibit bead binding. In contrast, the monoclonal Dsg1 antibody reduced bead binding also when applied for preincubation with beads, indicating a different mechanism underlying the reduction of Dsg1 adhesion ( n = 6 for each condition).

    Journal: Journal of Clinical Investigation

    Article Title: Pemphigus foliaceus IgG causes dissociation of desmoglein 1-containing junctions without blocking desmoglein 1 transinteraction

    doi: 10.1172/JCI23475

    Figure Lengend Snippet: Effect of PF-IgGs on binding of Dsg1-coated beads to HaCaT cells. Beads were allowed to settle on the surface of HaCaT cells for 30 minutes (control). Number of bound beads was reduced by simultaneous incubation of EGTA (5 mM, 30 minutes). Incubation of monolayers with attached beads for an additional 30 minutes with IgG fractions from 2 patients (PF1- and PF2-IgG, 35 μg/ml each) as well as with a monoclonal antibody (1:50) directed against the extracellular domain of human Dsg1 significantly reduced the number of bound beads (label in white box and white bars). Immunoabsorption using Dsg1-coated beads but not control absorption using VE-cadherin–labeled beads completely abolished the effect of PF-IgGs on bead adhesion. Preincubation of HaCaT cells with PF-IgGs prior to bead settlement also reduced bead binding (label in gray box and gray bars) whereas preincubation of beads with PF-IgGs did not inhibit bead binding. In contrast, the monoclonal Dsg1 antibody reduced bead binding also when applied for preincubation with beads, indicating a different mechanism underlying the reduction of Dsg1 adhesion ( n = 6 for each condition).

    Article Snippet: Negative controls were performed using Dsg1-coated beads incubated on the surface of HaCaT cells in the presence of 5 mM EGTA (Roth) or in the presence of monoclonal mouse IgG antibody (1:50 in HBSS) directed against Dsg-1 (clone p124; Progen Industries Ltd.).

    Techniques: Binding Assay, Incubation, Labeling

    ATP's antimicrobial effect is attributable to its iron-chelating activity. (a) Blocking effects of MgCl 2 and FeCl 3 on ATP's anti- M. intracellulare antimicrobial activity. (b) Anti- M. intracellulare activities of ATP and various metal-chelating agents (pyrophosphate (PPi), EDTA, EGTA) and blocking effects of MgCl 2 . (c) Anti- S. aureus activities of ATP and various metal-chelating agents. (d) Ferric ion-chelating activities of ATP and EDTA measured by CAS assay. (e) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against M. intracellulare . (f) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against P. aeruginosa . (g) Siderophore production by ATP-resistant E. coli during the course of cultivation. (h) Siderophore production by two ATP-resistant K. pneumoniae strains during 48-h cultivation in the presence of ATP. (i) Siderophore production by ATP-susceptible S. aureus and ATP-resistant E. coli during 24-h cultivation in the presence of ATP (5 mM). (j) Siderophore production by ATP-susceptible and ATP-resistant strains of P. aeruginosa during 24-h cultivation.

    Journal: Scientific Reports

    Article Title: ATP Exhibits Antimicrobial Action by Inhibiting Bacterial Utilization of Ferric Ions

    doi: 10.1038/srep08610

    Figure Lengend Snippet: ATP's antimicrobial effect is attributable to its iron-chelating activity. (a) Blocking effects of MgCl 2 and FeCl 3 on ATP's anti- M. intracellulare antimicrobial activity. (b) Anti- M. intracellulare activities of ATP and various metal-chelating agents (pyrophosphate (PPi), EDTA, EGTA) and blocking effects of MgCl 2 . (c) Anti- S. aureus activities of ATP and various metal-chelating agents. (d) Ferric ion-chelating activities of ATP and EDTA measured by CAS assay. (e) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against M. intracellulare . (f) Ferric ion-mediated reduction of antimicrobial effects of ATP and EDTA against P. aeruginosa . (g) Siderophore production by ATP-resistant E. coli during the course of cultivation. (h) Siderophore production by two ATP-resistant K. pneumoniae strains during 48-h cultivation in the presence of ATP. (i) Siderophore production by ATP-susceptible S. aureus and ATP-resistant E. coli during 24-h cultivation in the presence of ATP (5 mM). (j) Siderophore production by ATP-susceptible and ATP-resistant strains of P. aeruginosa during 24-h cultivation.

    Article Snippet: Special agents The following agents were used: ATP (Sigma Aldrich Co., St. Louis, MO; MP Biomedicals, Solon, OH, USA, Calbiochem Co., La Jolla, CA, and Roche Diagnostic Co., Indianapolis, IN), ADP (Sigma), AMP (Sigma), adenosine (Sigma), benzoylbenzoyl ATP (Sigma), oxidized ATP (Sigma), suramin (Wako, Tokyo, Japan), MIA (methyl isobutyl amiloride, Wako), DIDS (4,4′- Diisothiocyanatostilbene-2,2′-disulfonic acid, Sigma), EDTA (Dojindo, Tokyo, Japan), EGTA (Dojindo), dipyridyl (Sigma), CAS (Chrome Azurol S, Sigma), E. coli S17-1 and pK18mobSacB (kindly provided by Dr. F. Taguchi, Okayama University), Instagene matrix (Bio-Rad, Hercules, CA), KOD-Plus (Toyobo, Osaka, Japan), Wizard SV Gel and PCR cleanup system (Promeg, Madison, WI), Ligation High (Toyobo), Protein Assay Rapid Kit (Wako), [14 C] isoleucine (Moravek Biochemicals, Inc, Brea, CA), [14 C]uracil (Moravek Biochemicals, Inc.), α-defensin-1 (Peptide institute, Inc, Osaka, Japan.), cathepsin G (Sigma), vancomycin (Wako), clarithromycin (Taisho-Toyama Pharmaceutical Co., Tokyo), rifampin (Daiichi Sankyo Co., Tokyo), and ethambutol (Sigma), gatifloxacin (Wako), FLUOS (Sigma), LB medium (Invitrogen, San Diego, CA), M9 medium (prepared by our laboratory), Heart infusion agar (Eiken Chemical Co., Tokyo, Japan), Middlebrook 7H9 medium (Becton Dickinson, Cockeysville, MD), and Middlebrook 7H11 medium (Becton Dickinson).

    Techniques: Activity Assay, Blocking Assay

    Typical force versus relative deformation profiles for MDA-MB-468 cells labeled with various dyes: a control cell (1, red), a EGTA treated cell (2, gray), and cells labeled with 5 μM CFDA-SE (3, blue), CMFDA (4, green), CMTMR (5, orange) and Calcein

    Journal: The journal of physical chemistry. B

    Article Title: Cell tracing dyes significantly change single cell mechanics

    doi: 10.1021/jp8103358

    Figure Lengend Snippet: Typical force versus relative deformation profiles for MDA-MB-468 cells labeled with various dyes: a control cell (1, red), a EGTA treated cell (2, gray), and cells labeled with 5 μM CFDA-SE (3, blue), CMFDA (4, green), CMTMR (5, orange) and Calcein

    Article Snippet: To investigate role of calcium ions in cell stiffening MDA-MB-468 cells were incubated with 5 mM ethylene glycol tetraacetic acid, EGTA (Fisher Scientific, Fairlawn, NJ) for 30 min before compression.

    Techniques: Multiple Displacement Amplification, Labeling

    Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L BAPTA-AM + 0.5 mmol/L EGTA, n = 5) or introducing a

    Journal: Neuroscience Bulletin

    Article Title: Biophotons Contribute to Retinal Dark Noise

    doi: 10.1007/s12264-016-0029-6

    Figure Lengend Snippet: Effects of Ca 2+ and PDE on biophotonic activity in rat retina. A – D Removing intra- and extracellular Ca 2+ together ( A and B , 10 µmol/L BAPTA-AM + 0.5 mmol/L EGTA, n = 5) or introducing a

    Article Snippet: A phosphodiesterase 6 (PDE6) inhibitor (Zaprinast, 100 nmol/L, Sigma, St. Louis, MO), BAPTA-AM (10 μmol/L, Molecular Probes, Eugene, OR), and EGTA (0.5 mmol/L, Amresco, Solon, OH) were initially dissolved in DMSO and then diluted to their final concentrations in ACSF or Ringer’s solution.

    Techniques: Activity Assay

    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM EGTA or 10 μM BAPTA-AM, for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Neuroprotection and spatial memory enhancement of four herbal mixture extract in HT22 hippocampal cells and a mouse model of focal cerebral ischemia

    doi: 10.1186/s12906-015-0741-1

    Figure Lengend Snippet: Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM EGTA or 10 μM BAPTA-AM, for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P

    Article Snippet: BAPTA-AM and EGTA were purchased from Tocris Bioscience (Ellisville, MO, USA).

    Techniques: Fluorescence

    Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and EGTA or EDTA was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the

    Journal: Infection and Immunity

    Article Title: Listeriolysin O-Dependent Bacterial Entry into the Cytoplasm Is Required for Calpain Activation and Interleukin-1? Secretion in Macrophages Infected with Listeria monocytogenes ▿

    doi: 10.1128/IAI.01143-09

    Figure Lengend Snippet: Involvement of intracellular calcium elevation in IL-1α secretion. Macrophages were infected with L. monocytogenes for 1 h, and EGTA or EDTA was added to the cultures at 3 h after infection (A). Alternatively, macrophages were infected with the

    Article Snippet: EDTA and EGTA were purchased from Nacalai Tesque Inc. (Kyoto, Japan).

    Techniques: Infection

    Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM EGTA, 5 mM Ni 2+ , 125 μM verapamil (VPL), or control HBSS. Results are means ± SEM for four to six experiments. ∗, P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Interactions between HMR 3647, a New Ketolide, and Human Polymorphonuclear Neutrophils

    doi:

    Figure Lengend Snippet: Effect of Ca 2+ chelation and Ca 2+ channel inhibitors on HMR 3647 uptake. (A) PMNs were incubated for 5 to 60 min in the presence of 1 mM EGTA, 5 mM Ni 2+ , 125 μM verapamil (VPL), or control HBSS. Results are means ± SEM for four to six experiments. ∗, P

    Article Snippet: The uptake kinetics of macrolides was assessed first in Ca2+ -depleted HBSS (Gibco), supplemented with 1 mM EGTA (Merck), 1 mM magnesium chloride (Merck), and 4.2 mM sodium bicarbonate (NaHCO3 ) (Diagnostic Pasteur).

    Techniques: Incubation

    Excessive Ca 2+ -Independent Actomyosin Associations during Diastole Promote Enhanced Myocyte Shortening and Incomplete Relaxation of Hand > Act57B A295S Cardiomyocytes (A) M modes generated from the same region of a three-week-old Hand > Act57B WT heart reveal graded responses of cardiac diameters following exposure to distinct small-molecule compounds. Red arrowheads indicate the position of the heart wall edges during diastole, upon incubation with EGTA/EGTA,AM, and finally, following the addition of blebbistatin. Incubation with EGTA/EGTA,AM resulted in complete cessation of wall motion. Relative to the diameter during diastole, each treatment induced a slight increase in diameter across the heart tube. (B) Significant, incremental increases in cardiac diameters were verified for all genotypes following extra- and intracellular Ca 2+ chelation and upon blebbistatin incubation. (C) The average change in diameter across the heart wall in response to EGTA/EGTA,AM was similar among all lines. (D) Blebbistatin treatment prompted a significantly greater response across the wall of Hand > Act57B A295S hearts relative to that observed for Hand × yw and Hand > Act57B WT hearts. Data are presented as mean ± SEM (n = 21). ***p ≤ 0.001. See also Figure S3 .

    Journal: Cell reports

    Article Title: Distortion of the Actin A-Triad Results in Contractile Disinhibition and Cardiomyopathy

    doi: 10.1016/j.celrep.2017.08.070

    Figure Lengend Snippet: Excessive Ca 2+ -Independent Actomyosin Associations during Diastole Promote Enhanced Myocyte Shortening and Incomplete Relaxation of Hand > Act57B A295S Cardiomyocytes (A) M modes generated from the same region of a three-week-old Hand > Act57B WT heart reveal graded responses of cardiac diameters following exposure to distinct small-molecule compounds. Red arrowheads indicate the position of the heart wall edges during diastole, upon incubation with EGTA/EGTA,AM, and finally, following the addition of blebbistatin. Incubation with EGTA/EGTA,AM resulted in complete cessation of wall motion. Relative to the diameter during diastole, each treatment induced a slight increase in diameter across the heart tube. (B) Significant, incremental increases in cardiac diameters were verified for all genotypes following extra- and intracellular Ca 2+ chelation and upon blebbistatin incubation. (C) The average change in diameter across the heart wall in response to EGTA/EGTA,AM was similar among all lines. (D) Blebbistatin treatment prompted a significantly greater response across the wall of Hand > Act57B A295S hearts relative to that observed for Hand × yw and Hand > Act57B WT hearts. Data are presented as mean ± SEM (n = 21). ***p ≤ 0.001. See also Figure S3 .

    Article Snippet: After initial filming, the AH was replaced with AH containing 10 mM EGTA and 100 μM EGTA,AM (AAT Bioquest).

    Techniques: Generated, Incubation