egta Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher egta
    Modeling VGCC-dependent glutamate miniature release. ( a ) Schematics illustrating VCell simulations. As in Fig. 6c the black dot indicate space taken up by the vesicle in the XY plane 2.5 nm above the active zone; gray circle, vesicle projection on the active zone plane; green dots, assumed positions of Ca 2+ -release sensors, grid 5 nm. ( b ) Examples of average [ Ca 2+ ] transients at release sensors produced by single VGCC openings (for 0.33 ms) for 4 different VGCC-release sensor distances. Insert, corresponding vesicle fusion probabilities. ( c ) Color-coded map showing dependency of vesicle fusion probability p v (Δ t,d ) on VGCC-vesicle distance d and VGCC open-channel duration Δ t . ( d ) Frequency histograms φ (Δ t ) for the durations of spontaneous P/Q-, N-, and R-type channel opening at V rest obtained using the VGCC gating model 12 ( Fig. 5a ). ( e ) Dependencies of vesicle fusion probability p v ( d ) on VGCC-vesicle distance for different VGCC subtypes. ( f ) Frequency histogram ψ ( d ) for the relative VGCC-vesicle distances in the Clustered model (n = 60 simulated active zones). ( g ) Dependency of spontaneous P/Q-, N-, and R-type channel opening on V rest , calculated using the six-state VGCC gating model 12 ( Fig. 5a ). ( h ) Distribution of V rest in cultured hippocampal neurons (mean 71.9 ± 0.7 mV, n = 98 neurons). ( i ) Cumulative fractions of VGCC-mediated mEPSCs and VGCC numbers plotted as functions of the distance from the vesicular release sensor. ~90% of all VGCC-dependent minis are mediated by only ~20% of all VGCCs present in the active zone located within 70 nm of docked vesicles. ( j ) Model predictions for the effects of <t>BAPTA</t> and <t>EGTA</t> on VGCC-dependent mEPSC frequency. Dotted lines, experimental effects of BAPTA-AM and EGTA-AM as estimated in Fig. 4e .
    Egta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4669 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Thermo Fisher
    Average 99 stars, based on 4669 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    Millipore ethylene glycol tetraacetic acid
    Modeling VGCC-dependent glutamate miniature release. ( a ) Schematics illustrating VCell simulations. As in Fig. 6c the black dot indicate space taken up by the vesicle in the XY plane 2.5 nm above the active zone; gray circle, vesicle projection on the active zone plane; green dots, assumed positions of Ca 2+ -release sensors, grid 5 nm. ( b ) Examples of average [ Ca 2+ ] transients at release sensors produced by single VGCC openings (for 0.33 ms) for 4 different VGCC-release sensor distances. Insert, corresponding vesicle fusion probabilities. ( c ) Color-coded map showing dependency of vesicle fusion probability p v (Δ t,d ) on VGCC-vesicle distance d and VGCC open-channel duration Δ t . ( d ) Frequency histograms φ (Δ t ) for the durations of spontaneous P/Q-, N-, and R-type channel opening at V rest obtained using the VGCC gating model 12 ( Fig. 5a ). ( e ) Dependencies of vesicle fusion probability p v ( d ) on VGCC-vesicle distance for different VGCC subtypes. ( f ) Frequency histogram ψ ( d ) for the relative VGCC-vesicle distances in the Clustered model (n = 60 simulated active zones). ( g ) Dependency of spontaneous P/Q-, N-, and R-type channel opening on V rest , calculated using the six-state VGCC gating model 12 ( Fig. 5a ). ( h ) Distribution of V rest in cultured hippocampal neurons (mean 71.9 ± 0.7 mV, n = 98 neurons). ( i ) Cumulative fractions of VGCC-mediated mEPSCs and VGCC numbers plotted as functions of the distance from the vesicular release sensor. ~90% of all VGCC-dependent minis are mediated by only ~20% of all VGCCs present in the active zone located within 70 nm of docked vesicles. ( j ) Model predictions for the effects of <t>BAPTA</t> and <t>EGTA</t> on VGCC-dependent mEPSC frequency. Dotted lines, experimental effects of BAPTA-AM and EGTA-AM as estimated in Fig. 4e .
    Ethylene Glycol Tetraacetic Acid, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 653 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ethylene glycol tetraacetic acid/product/Millipore
    Average 99 stars, based on 653 article reviews
    Price from $9.99 to $1999.99
    ethylene glycol tetraacetic acid - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    Millipore ethylene glycol tetraacetic acid egta
    Homocysteine induced NETosis is ROS and Ca2 + influx dependent. Freshly isolated peripheral neutrophils were pretreated with either N-acetyl cysteine (1 mM) ( a ) or <t>Diphenyleneiodonium</t> (20 μM) ( b ) or BAPTA (10 μM) ( e ) or <t>EGTA</t> (10 mM) ( f ) or TMB-8 (20 μM) ( g ) for 30 minutes and followed by activation either in presence or absence of homocysteine alone ( a ) or homocysteine (100 μM) IL-6 (25 ng/ml), PMA (20 ng/ml), LPS (2 μg/ml) for three hours. Extent of NETs formation was measured after staining with SYTOX Green (5 μM) in fluorimeter. Data is represented as percentage of maximal SYTOX Green fluorescence. Statistically significant modulation in NETosis is denoted by asterisk ***p
    Ethylene Glycol Tetraacetic Acid Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 799 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ethylene glycol tetraacetic acid egta/product/Millipore
    Average 99 stars, based on 799 article reviews
    Price from $9.99 to $1999.99
    ethylene glycol tetraacetic acid egta - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    94
    Millipore egta am
    Homocysteine induced NETosis is ROS and Ca2 + influx dependent. Freshly isolated peripheral neutrophils were pretreated with either N-acetyl cysteine (1 mM) ( a ) or <t>Diphenyleneiodonium</t> (20 μM) ( b ) or BAPTA (10 μM) ( e ) or <t>EGTA</t> (10 mM) ( f ) or TMB-8 (20 μM) ( g ) for 30 minutes and followed by activation either in presence or absence of homocysteine alone ( a ) or homocysteine (100 μM) IL-6 (25 ng/ml), PMA (20 ng/ml), LPS (2 μg/ml) for three hours. Extent of NETs formation was measured after staining with SYTOX Green (5 μM) in fluorimeter. Data is represented as percentage of maximal SYTOX Green fluorescence. Statistically significant modulation in NETosis is denoted by asterisk ***p
    Egta Am, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 210 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta am/product/Millipore
    Average 94 stars, based on 210 article reviews
    Price from $9.99 to $1999.99
    egta am - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    egta  (Tocris)
    99
    Tocris egta
    Homocysteine induced NETosis is ROS and Ca2 + influx dependent. Freshly isolated peripheral neutrophils were pretreated with either N-acetyl cysteine (1 mM) ( a ) or <t>Diphenyleneiodonium</t> (20 μM) ( b ) or BAPTA (10 μM) ( e ) or <t>EGTA</t> (10 mM) ( f ) or TMB-8 (20 μM) ( g ) for 30 minutes and followed by activation either in presence or absence of homocysteine alone ( a ) or homocysteine (100 μM) IL-6 (25 ng/ml), PMA (20 ng/ml), LPS (2 μg/ml) for three hours. Extent of NETs formation was measured after staining with SYTOX Green (5 μM) in fluorimeter. Data is represented as percentage of maximal SYTOX Green fluorescence. Statistically significant modulation in NETosis is denoted by asterisk ***p
    Egta, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 275 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Tocris
    Average 99 stars, based on 275 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    92
    AnaSpec egta am
    CAX regulates cell substrate attachment. (a and b) Effect of CAX knockdown on chemotaxis. (a) Tracks of individual explants migrating toward beads soaked in Sdf-1 over the entire period of recording (left) or the initial 100 min (right). (b) Summary data (from three knockdowns) quantifying forward motion index, persistence, and velocity of migration. (c–g) Effect of CAX knockdown on cell dispersion. (c) Summary data from three knockdowns quantifying the size of control and morphant explants 30 min after plating. (d) Representative color-coded triangulation diagrams at time 0 and 6 h from control or morphant explants. (e) Time courses (from three knockdowns) quantifying triangle area. Data are normalized to control explants at time 0. (f) Transmitted light micrographs showing spreading of control or morphant explants over a 20-min period. Panels on the right show an overlay of the area occupied by the explants where red represents the extent of spreading. Bar, 500 µm. (g) Summary data from three knockdowns quantifying spread area. (h and i) Effect of Ca 2+ buffering on cell dispersion. Triangulation (h) and spreading (i) analysis of explants treated with cell-permeable Ca 2+ chelators (50 µM <t>BAPTA/EGTA-AM)</t> or DMSO 30 min before imaging. (j and k) Live cell imaging of CAX. Stills from time-lapse confocal imaging of explants coexpressing mRFP-CAX and either membrane GFP (j; bar, 10 µm) or focal adhesion kinase–GFP (k). Arrowheads mark small CAX-positive vesicles. (k) Bars: (top) 4 µm; (bottom) 1 µm. Heat map summarizes the mean projection for CAX-positive vesicles relative to the centroid of focal adhesions (intersection of white lines) within 5 × 5–µm regions of interest. Data are from 28 focal adhesions from two expression experiments. (l–n) Effect of CAX knockdown on focal adhesions. (l) Expression of focal adhesion kinase–GFP (left) and immunocytochemistry analysis using an antibody to phosphopaxillin (middle) in control and morphant explants. Overlays (right) show costaining of phalloidin. Bar, 20 µm. (m) Summary data (94–422 cells from two to three knockdowns) quantifying the number and size of labeled structures. (n) Lifetime analysis of focal adhesion kinase–GFP in control and morphant explants (30–58 focal adhesions from two knockdowns). 10 ng AMO1 was used for all knockdowns. Error bars represent SEM. *, P
    Egta Am, supplied by AnaSpec, used in various techniques. Bioz Stars score: 92/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta am/product/AnaSpec
    Average 92 stars, based on 99 article reviews
    Price from $9.99 to $1999.99
    egta am - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    92
    Boston BioProducts egta
    CAX regulates cell substrate attachment. (a and b) Effect of CAX knockdown on chemotaxis. (a) Tracks of individual explants migrating toward beads soaked in Sdf-1 over the entire period of recording (left) or the initial 100 min (right). (b) Summary data (from three knockdowns) quantifying forward motion index, persistence, and velocity of migration. (c–g) Effect of CAX knockdown on cell dispersion. (c) Summary data from three knockdowns quantifying the size of control and morphant explants 30 min after plating. (d) Representative color-coded triangulation diagrams at time 0 and 6 h from control or morphant explants. (e) Time courses (from three knockdowns) quantifying triangle area. Data are normalized to control explants at time 0. (f) Transmitted light micrographs showing spreading of control or morphant explants over a 20-min period. Panels on the right show an overlay of the area occupied by the explants where red represents the extent of spreading. Bar, 500 µm. (g) Summary data from three knockdowns quantifying spread area. (h and i) Effect of Ca 2+ buffering on cell dispersion. Triangulation (h) and spreading (i) analysis of explants treated with cell-permeable Ca 2+ chelators (50 µM <t>BAPTA/EGTA-AM)</t> or DMSO 30 min before imaging. (j and k) Live cell imaging of CAX. Stills from time-lapse confocal imaging of explants coexpressing mRFP-CAX and either membrane GFP (j; bar, 10 µm) or focal adhesion kinase–GFP (k). Arrowheads mark small CAX-positive vesicles. (k) Bars: (top) 4 µm; (bottom) 1 µm. Heat map summarizes the mean projection for CAX-positive vesicles relative to the centroid of focal adhesions (intersection of white lines) within 5 × 5–µm regions of interest. Data are from 28 focal adhesions from two expression experiments. (l–n) Effect of CAX knockdown on focal adhesions. (l) Expression of focal adhesion kinase–GFP (left) and immunocytochemistry analysis using an antibody to phosphopaxillin (middle) in control and morphant explants. Overlays (right) show costaining of phalloidin. Bar, 20 µm. (m) Summary data (94–422 cells from two to three knockdowns) quantifying the number and size of labeled structures. (n) Lifetime analysis of focal adhesion kinase–GFP in control and morphant explants (30–58 focal adhesions from two knockdowns). 10 ng AMO1 was used for all knockdowns. Error bars represent SEM. *, P
    Egta, supplied by Boston BioProducts, used in various techniques. Bioz Stars score: 92/100, based on 177 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Boston BioProducts
    Average 92 stars, based on 177 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    95
    Thermo Fisher np egta am
    CAX regulates cell substrate attachment. (a and b) Effect of CAX knockdown on chemotaxis. (a) Tracks of individual explants migrating toward beads soaked in Sdf-1 over the entire period of recording (left) or the initial 100 min (right). (b) Summary data (from three knockdowns) quantifying forward motion index, persistence, and velocity of migration. (c–g) Effect of CAX knockdown on cell dispersion. (c) Summary data from three knockdowns quantifying the size of control and morphant explants 30 min after plating. (d) Representative color-coded triangulation diagrams at time 0 and 6 h from control or morphant explants. (e) Time courses (from three knockdowns) quantifying triangle area. Data are normalized to control explants at time 0. (f) Transmitted light micrographs showing spreading of control or morphant explants over a 20-min period. Panels on the right show an overlay of the area occupied by the explants where red represents the extent of spreading. Bar, 500 µm. (g) Summary data from three knockdowns quantifying spread area. (h and i) Effect of Ca 2+ buffering on cell dispersion. Triangulation (h) and spreading (i) analysis of explants treated with cell-permeable Ca 2+ chelators (50 µM <t>BAPTA/EGTA-AM)</t> or DMSO 30 min before imaging. (j and k) Live cell imaging of CAX. Stills from time-lapse confocal imaging of explants coexpressing mRFP-CAX and either membrane GFP (j; bar, 10 µm) or focal adhesion kinase–GFP (k). Arrowheads mark small CAX-positive vesicles. (k) Bars: (top) 4 µm; (bottom) 1 µm. Heat map summarizes the mean projection for CAX-positive vesicles relative to the centroid of focal adhesions (intersection of white lines) within 5 × 5–µm regions of interest. Data are from 28 focal adhesions from two expression experiments. (l–n) Effect of CAX knockdown on focal adhesions. (l) Expression of focal adhesion kinase–GFP (left) and immunocytochemistry analysis using an antibody to phosphopaxillin (middle) in control and morphant explants. Overlays (right) show costaining of phalloidin. Bar, 20 µm. (m) Summary data (94–422 cells from two to three knockdowns) quantifying the number and size of labeled structures. (n) Lifetime analysis of focal adhesion kinase–GFP in control and morphant explants (30–58 focal adhesions from two knockdowns). 10 ng AMO1 was used for all knockdowns. Error bars represent SEM. *, P
    Np Egta Am, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 78 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/np egta am/product/Thermo Fisher
    Average 95 stars, based on 78 article reviews
    Price from $9.99 to $1999.99
    np egta am - by Bioz Stars, 2020-10
    95/100 stars
      Buy from Supplier

    93
    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: 93/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Dojindo Labs
    Average 93 stars, based on 106 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    93
    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: 93/100, based on 299 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Fisher Scientific
    Average 93 stars, based on 299 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    92
    Merck & Co egta
    Protoplast shrinkage at <t>55°C</t> HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM <t>EGTA</t> prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p
    Egta, supplied by Merck & Co, used in various techniques. Bioz Stars score: 92/100, based on 233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Merck & Co
    Average 92 stars, based on 233 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher egta tetra acetoxymethyl ester egta am
    Protoplast shrinkage at <t>55°C</t> HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM <t>EGTA</t> prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p
    Egta Tetra Acetoxymethyl Ester Egta Am, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta tetra acetoxymethyl ester egta am/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    egta tetra acetoxymethyl ester egta am - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    Millipore ethylene glycol bis beta aminoethyl ether n n n n tetraacetic acid tetrasodium salt
    Protoplast shrinkage at <t>55°C</t> HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM <t>EGTA</t> prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p
    Ethylene Glycol Bis Beta Aminoethyl Ether N N N N Tetraacetic Acid Tetrasodium Salt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ethylene glycol bis beta aminoethyl ether n n n n tetraacetic acid tetrasodium salt/product/Millipore
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    ethylene glycol bis beta aminoethyl ether n n n n tetraacetic acid tetrasodium salt - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    99
    Millipore egta ethylene glycol bis β aminoethyl ether n
    Protoplast shrinkage at <t>55°C</t> HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM <t>EGTA</t> prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p
    Egta Ethylene Glycol Bis β Aminoethyl Ether N, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta ethylene glycol bis β aminoethyl ether n/product/Millipore
    Average 99 stars, based on 38 article reviews
    Price from $9.99 to $1999.99
    egta ethylene glycol bis β aminoethyl ether n - by Bioz Stars, 2020-10
    99/100 stars
      Buy from Supplier

    92
    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: 92/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Carl Roth GmbH
    Average 92 stars, based on 120 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    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 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Amresco
    Average 93 stars, based on 83 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    93/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology 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 Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1043 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1043 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-10
    94/100 stars
      Buy from Supplier

    Image Search Results


    Modeling VGCC-dependent glutamate miniature release. ( a ) Schematics illustrating VCell simulations. As in Fig. 6c the black dot indicate space taken up by the vesicle in the XY plane 2.5 nm above the active zone; gray circle, vesicle projection on the active zone plane; green dots, assumed positions of Ca 2+ -release sensors, grid 5 nm. ( b ) Examples of average [ Ca 2+ ] transients at release sensors produced by single VGCC openings (for 0.33 ms) for 4 different VGCC-release sensor distances. Insert, corresponding vesicle fusion probabilities. ( c ) Color-coded map showing dependency of vesicle fusion probability p v (Δ t,d ) on VGCC-vesicle distance d and VGCC open-channel duration Δ t . ( d ) Frequency histograms φ (Δ t ) for the durations of spontaneous P/Q-, N-, and R-type channel opening at V rest obtained using the VGCC gating model 12 ( Fig. 5a ). ( e ) Dependencies of vesicle fusion probability p v ( d ) on VGCC-vesicle distance for different VGCC subtypes. ( f ) Frequency histogram ψ ( d ) for the relative VGCC-vesicle distances in the Clustered model (n = 60 simulated active zones). ( g ) Dependency of spontaneous P/Q-, N-, and R-type channel opening on V rest , calculated using the six-state VGCC gating model 12 ( Fig. 5a ). ( h ) Distribution of V rest in cultured hippocampal neurons (mean 71.9 ± 0.7 mV, n = 98 neurons). ( i ) Cumulative fractions of VGCC-mediated mEPSCs and VGCC numbers plotted as functions of the distance from the vesicular release sensor. ~90% of all VGCC-dependent minis are mediated by only ~20% of all VGCCs present in the active zone located within 70 nm of docked vesicles. ( j ) Model predictions for the effects of BAPTA and EGTA on VGCC-dependent mEPSC frequency. Dotted lines, experimental effects of BAPTA-AM and EGTA-AM as estimated in Fig. 4e .

    Journal: Nature neuroscience

    Article Title: Differential triggering of spontaneous glutamate release by P/Q-, N-, and R-type Ca2+ channels

    doi: 10.1038/nn.3563

    Figure Lengend Snippet: Modeling VGCC-dependent glutamate miniature release. ( a ) Schematics illustrating VCell simulations. As in Fig. 6c the black dot indicate space taken up by the vesicle in the XY plane 2.5 nm above the active zone; gray circle, vesicle projection on the active zone plane; green dots, assumed positions of Ca 2+ -release sensors, grid 5 nm. ( b ) Examples of average [ Ca 2+ ] transients at release sensors produced by single VGCC openings (for 0.33 ms) for 4 different VGCC-release sensor distances. Insert, corresponding vesicle fusion probabilities. ( c ) Color-coded map showing dependency of vesicle fusion probability p v (Δ t,d ) on VGCC-vesicle distance d and VGCC open-channel duration Δ t . ( d ) Frequency histograms φ (Δ t ) for the durations of spontaneous P/Q-, N-, and R-type channel opening at V rest obtained using the VGCC gating model 12 ( Fig. 5a ). ( e ) Dependencies of vesicle fusion probability p v ( d ) on VGCC-vesicle distance for different VGCC subtypes. ( f ) Frequency histogram ψ ( d ) for the relative VGCC-vesicle distances in the Clustered model (n = 60 simulated active zones). ( g ) Dependency of spontaneous P/Q-, N-, and R-type channel opening on V rest , calculated using the six-state VGCC gating model 12 ( Fig. 5a ). ( h ) Distribution of V rest in cultured hippocampal neurons (mean 71.9 ± 0.7 mV, n = 98 neurons). ( i ) Cumulative fractions of VGCC-mediated mEPSCs and VGCC numbers plotted as functions of the distance from the vesicular release sensor. ~90% of all VGCC-dependent minis are mediated by only ~20% of all VGCCs present in the active zone located within 70 nm of docked vesicles. ( j ) Model predictions for the effects of BAPTA and EGTA on VGCC-dependent mEPSC frequency. Dotted lines, experimental effects of BAPTA-AM and EGTA-AM as estimated in Fig. 4e .

    Article Snippet: Drugs were directly added to the recirculating perfusion system (total volume 10 ml) to achieve the following final concentrations: 0.25 μM ω-Aga (Bachem, Germany), 5 μM ω-Ctx, 0.5 μM SNX 482 (Abcam, UK), 1.0 μM TTA-P2 (Merck, USA), 100 μM CdCl2 (Sigma), 20 μM EGTA-AM and 20 μM BAPTA-AM (Invitrogen, USA).

    Techniques: Produced, Mass Spectrometry, Cell Culture

    Differential effects of slow (EGTA) and fast (BAPTA) exogenous Ca 2+ buffers on VGCC-dependent minis. ( a, b ) Time-course of mEPSC frequency changes during incubation in BAPTA-AM ( a ) and EGTA-AM ( b ). Left, mEPSC traces from representative experiments before and after addition of the Ca 2+ chelators. Right, average responses in N = 7 cells for BAPTA-AM and N = 9 cells for EGTA-AM. ( c, d ) Differential effects of VGCC blockers on mEPSC frequency in BAPTA-AM ( c ) and in EGTA-AM ( d ) pre-treated cultures (both in ( c ) and ( d ) N = 7 cells for ω-Aga and ω-Ctx, and N = 6 cells for ω-Aga, ω-Ctx, and SNX). To determine the remaining fraction of mEPSCs sensitive to VGCC blockade after BAPTA-AM and EGTA-AM treatment (shown on the right), the initial mEPSC frequencies in this set of experiments were normalized to the effects of BAPTA-AM or EGTA-AM determined in ( a ) and ( b ). In contrast to EGTA-AM, pretreatment with BAPTA-AM almost completely occluded the effect of toxins on miniature release. All data are mean ± s.e.m, *** P

    Journal: Nature neuroscience

    Article Title: Differential triggering of spontaneous glutamate release by P/Q-, N-, and R-type Ca2+ channels

    doi: 10.1038/nn.3563

    Figure Lengend Snippet: Differential effects of slow (EGTA) and fast (BAPTA) exogenous Ca 2+ buffers on VGCC-dependent minis. ( a, b ) Time-course of mEPSC frequency changes during incubation in BAPTA-AM ( a ) and EGTA-AM ( b ). Left, mEPSC traces from representative experiments before and after addition of the Ca 2+ chelators. Right, average responses in N = 7 cells for BAPTA-AM and N = 9 cells for EGTA-AM. ( c, d ) Differential effects of VGCC blockers on mEPSC frequency in BAPTA-AM ( c ) and in EGTA-AM ( d ) pre-treated cultures (both in ( c ) and ( d ) N = 7 cells for ω-Aga and ω-Ctx, and N = 6 cells for ω-Aga, ω-Ctx, and SNX). To determine the remaining fraction of mEPSCs sensitive to VGCC blockade after BAPTA-AM and EGTA-AM treatment (shown on the right), the initial mEPSC frequencies in this set of experiments were normalized to the effects of BAPTA-AM or EGTA-AM determined in ( a ) and ( b ). In contrast to EGTA-AM, pretreatment with BAPTA-AM almost completely occluded the effect of toxins on miniature release. All data are mean ± s.e.m, *** P

    Article Snippet: Drugs were directly added to the recirculating perfusion system (total volume 10 ml) to achieve the following final concentrations: 0.25 μM ω-Aga (Bachem, Germany), 5 μM ω-Ctx, 0.5 μM SNX 482 (Abcam, UK), 1.0 μM TTA-P2 (Merck, USA), 100 μM CdCl2 (Sigma), 20 μM EGTA-AM and 20 μM BAPTA-AM (Invitrogen, USA).

    Techniques: Incubation

    FM dye imaging of action potential-evoked exocytosis reveals similar sensitivity of evoked and VGCC-dependent miniature release to presynaptic Ca 2+ chelation. ( a ) Experimental paradigm (see also Online Methods). ( b ) Example of fluorescence loss in individual synaptic boutons during the time course of a typical control experiment. Arrows: individual boutons examined in ( c ). Scale bar 5 μm. ( c ) FM dye de-staining profiles in boutons 1 and 2; exponential fits of de-staining rates at rest ( k rest ) and during 0.5 Hz action potential stimulation ( k stim ) are shown by red dashed lines. The effective specific action potential-evoked FM dye (SRC1) de-staining rate in each bouton was calculated as k AP = k stim – k rest . ( d ) Effect of BAPTA-AM and EGTA-AM on SRC1 de-staining kinetics. Average de-staining profiles from three representative experiments: Control (black), BAPTA-AM (gray) and EGTA-AM (white) symbols. Each profile is an average of 100–200 boutons. ( e ) Comparison of BAPTA-AM and EGTA-AM relative effects on the rate of action potential-evoked vesicular exocytosis k AP (left 2 bars, BAPTA-AM N = 6 experiments, EGTA-AM N = 5 experiments) and on the frequency of VGCC-dependent minis (right 2 bars). The latter was calculated from electrophysiological mEPSC recordings in Figs. 1 and 3 as detailed in Online Methods. Data are mean ± s.e.m, * P

    Journal: Nature neuroscience

    Article Title: Differential triggering of spontaneous glutamate release by P/Q-, N-, and R-type Ca2+ channels

    doi: 10.1038/nn.3563

    Figure Lengend Snippet: FM dye imaging of action potential-evoked exocytosis reveals similar sensitivity of evoked and VGCC-dependent miniature release to presynaptic Ca 2+ chelation. ( a ) Experimental paradigm (see also Online Methods). ( b ) Example of fluorescence loss in individual synaptic boutons during the time course of a typical control experiment. Arrows: individual boutons examined in ( c ). Scale bar 5 μm. ( c ) FM dye de-staining profiles in boutons 1 and 2; exponential fits of de-staining rates at rest ( k rest ) and during 0.5 Hz action potential stimulation ( k stim ) are shown by red dashed lines. The effective specific action potential-evoked FM dye (SRC1) de-staining rate in each bouton was calculated as k AP = k stim – k rest . ( d ) Effect of BAPTA-AM and EGTA-AM on SRC1 de-staining kinetics. Average de-staining profiles from three representative experiments: Control (black), BAPTA-AM (gray) and EGTA-AM (white) symbols. Each profile is an average of 100–200 boutons. ( e ) Comparison of BAPTA-AM and EGTA-AM relative effects on the rate of action potential-evoked vesicular exocytosis k AP (left 2 bars, BAPTA-AM N = 6 experiments, EGTA-AM N = 5 experiments) and on the frequency of VGCC-dependent minis (right 2 bars). The latter was calculated from electrophysiological mEPSC recordings in Figs. 1 and 3 as detailed in Online Methods. Data are mean ± s.e.m, * P

    Article Snippet: Drugs were directly added to the recirculating perfusion system (total volume 10 ml) to achieve the following final concentrations: 0.25 μM ω-Aga (Bachem, Germany), 5 μM ω-Ctx, 0.5 μM SNX 482 (Abcam, UK), 1.0 μM TTA-P2 (Merck, USA), 100 μM CdCl2 (Sigma), 20 μM EGTA-AM and 20 μM BAPTA-AM (Invitrogen, USA).

    Techniques: Imaging, Fluorescence, Staining

    Modeling action potential-evoked release in small hippocampal synapses. ( a ) Allosteric model of Ca 2+ activation of vesicle fusion 19 . ( b ) Presynaptic bouton geometry used in VCell simulations. Scale bar 0.5 μm. ( c ) Representative distributions of VGCCs and vesicles in the active zone for Clustered (left) and Random (right) models. Top, XY cross-sections 2.5 nm above the active zone; bottom, XZ cross-sections corresponding to black dashed lines in the XY plane. Blue dashed lines, active zone borders; brown dots, VGCCs; black dots, space occupied by vesicles; gray circles, vesicle projections on the XY plane; green dots, locations of Ca 2+ -release sensors, grid 5 nm. ( d ) Simulation results corresponding to geometries in ( c ). Top, action potential waveform; middle, average [ Ca 2+ ] transients at Ca 2+ -release sensors; bottom, corresponding release rates; legends, resulting fusion probabilities p v . ( e ) Cumulative probability plots of p v for Clustered and Random models. (n = 28 vesicles from 7 simulated synapses for each model). ( f ) Cumulative probability plots showing the average number of VGCCs located within a given distance from the vesicular Ca 2+ -release sensors (n = 240 vesicles from 60 simulated synapses). ( g ) Model predictions for inhibition of evoked release by BAPTA and EGTA. Dotted lines show the experimental effects of BAPTA-AM and EGTA-AM as determined in Fig. 4e . ( h ) Dependency of p v on Δ[ Ca 2+ ] total simulated by progressive deletion of active VGCCs. Data are from 5 simulated synapses, each point represents average p v for 4 release-ready vesicles. Data on both axes are normalized to the corresponding maximal values at basal conditions. Dotted lines, fitted power function, with the slope corresponding to Ca 2+ current cooperativity m ICa = 2.46.

    Journal: Nature neuroscience

    Article Title: Differential triggering of spontaneous glutamate release by P/Q-, N-, and R-type Ca2+ channels

    doi: 10.1038/nn.3563

    Figure Lengend Snippet: Modeling action potential-evoked release in small hippocampal synapses. ( a ) Allosteric model of Ca 2+ activation of vesicle fusion 19 . ( b ) Presynaptic bouton geometry used in VCell simulations. Scale bar 0.5 μm. ( c ) Representative distributions of VGCCs and vesicles in the active zone for Clustered (left) and Random (right) models. Top, XY cross-sections 2.5 nm above the active zone; bottom, XZ cross-sections corresponding to black dashed lines in the XY plane. Blue dashed lines, active zone borders; brown dots, VGCCs; black dots, space occupied by vesicles; gray circles, vesicle projections on the XY plane; green dots, locations of Ca 2+ -release sensors, grid 5 nm. ( d ) Simulation results corresponding to geometries in ( c ). Top, action potential waveform; middle, average [ Ca 2+ ] transients at Ca 2+ -release sensors; bottom, corresponding release rates; legends, resulting fusion probabilities p v . ( e ) Cumulative probability plots of p v for Clustered and Random models. (n = 28 vesicles from 7 simulated synapses for each model). ( f ) Cumulative probability plots showing the average number of VGCCs located within a given distance from the vesicular Ca 2+ -release sensors (n = 240 vesicles from 60 simulated synapses). ( g ) Model predictions for inhibition of evoked release by BAPTA and EGTA. Dotted lines show the experimental effects of BAPTA-AM and EGTA-AM as determined in Fig. 4e . ( h ) Dependency of p v on Δ[ Ca 2+ ] total simulated by progressive deletion of active VGCCs. Data are from 5 simulated synapses, each point represents average p v for 4 release-ready vesicles. Data on both axes are normalized to the corresponding maximal values at basal conditions. Dotted lines, fitted power function, with the slope corresponding to Ca 2+ current cooperativity m ICa = 2.46.

    Article Snippet: Drugs were directly added to the recirculating perfusion system (total volume 10 ml) to achieve the following final concentrations: 0.25 μM ω-Aga (Bachem, Germany), 5 μM ω-Ctx, 0.5 μM SNX 482 (Abcam, UK), 1.0 μM TTA-P2 (Merck, USA), 100 μM CdCl2 (Sigma), 20 μM EGTA-AM and 20 μM BAPTA-AM (Invitrogen, USA).

    Techniques: Activation Assay, Inhibition

    Homocysteine induced NETosis is ROS and Ca2 + influx dependent. Freshly isolated peripheral neutrophils were pretreated with either N-acetyl cysteine (1 mM) ( a ) or Diphenyleneiodonium (20 μM) ( b ) or BAPTA (10 μM) ( e ) or EGTA (10 mM) ( f ) or TMB-8 (20 μM) ( g ) for 30 minutes and followed by activation either in presence or absence of homocysteine alone ( a ) or homocysteine (100 μM) IL-6 (25 ng/ml), PMA (20 ng/ml), LPS (2 μg/ml) for three hours. Extent of NETs formation was measured after staining with SYTOX Green (5 μM) in fluorimeter. Data is represented as percentage of maximal SYTOX Green fluorescence. Statistically significant modulation in NETosis is denoted by asterisk ***p

    Journal: Scientific Reports

    Article Title: Elevated homocysteine levels in type 2 diabetes induce constitutive neutrophil extracellular traps

    doi: 10.1038/srep36362

    Figure Lengend Snippet: Homocysteine induced NETosis is ROS and Ca2 + influx dependent. Freshly isolated peripheral neutrophils were pretreated with either N-acetyl cysteine (1 mM) ( a ) or Diphenyleneiodonium (20 μM) ( b ) or BAPTA (10 μM) ( e ) or EGTA (10 mM) ( f ) or TMB-8 (20 μM) ( g ) for 30 minutes and followed by activation either in presence or absence of homocysteine alone ( a ) or homocysteine (100 μM) IL-6 (25 ng/ml), PMA (20 ng/ml), LPS (2 μg/ml) for three hours. Extent of NETs formation was measured after staining with SYTOX Green (5 μM) in fluorimeter. Data is represented as percentage of maximal SYTOX Green fluorescence. Statistically significant modulation in NETosis is denoted by asterisk ***p

    Article Snippet: Diphenyleneiodonium (DPI), Ethylene glycol tetra acetic acid (EGTA), BAPTA-AM (Sigma Chemicals, USA) was used to assess role of NADPH oxidase and Ca2+ influx on NETosis respectively.

    Techniques: Isolation, Activation Assay, Staining, Fluorescence

    CAX regulates cell substrate attachment. (a and b) Effect of CAX knockdown on chemotaxis. (a) Tracks of individual explants migrating toward beads soaked in Sdf-1 over the entire period of recording (left) or the initial 100 min (right). (b) Summary data (from three knockdowns) quantifying forward motion index, persistence, and velocity of migration. (c–g) Effect of CAX knockdown on cell dispersion. (c) Summary data from three knockdowns quantifying the size of control and morphant explants 30 min after plating. (d) Representative color-coded triangulation diagrams at time 0 and 6 h from control or morphant explants. (e) Time courses (from three knockdowns) quantifying triangle area. Data are normalized to control explants at time 0. (f) Transmitted light micrographs showing spreading of control or morphant explants over a 20-min period. Panels on the right show an overlay of the area occupied by the explants where red represents the extent of spreading. Bar, 500 µm. (g) Summary data from three knockdowns quantifying spread area. (h and i) Effect of Ca 2+ buffering on cell dispersion. Triangulation (h) and spreading (i) analysis of explants treated with cell-permeable Ca 2+ chelators (50 µM BAPTA/EGTA-AM) or DMSO 30 min before imaging. (j and k) Live cell imaging of CAX. Stills from time-lapse confocal imaging of explants coexpressing mRFP-CAX and either membrane GFP (j; bar, 10 µm) or focal adhesion kinase–GFP (k). Arrowheads mark small CAX-positive vesicles. (k) Bars: (top) 4 µm; (bottom) 1 µm. Heat map summarizes the mean projection for CAX-positive vesicles relative to the centroid of focal adhesions (intersection of white lines) within 5 × 5–µm regions of interest. Data are from 28 focal adhesions from two expression experiments. (l–n) Effect of CAX knockdown on focal adhesions. (l) Expression of focal adhesion kinase–GFP (left) and immunocytochemistry analysis using an antibody to phosphopaxillin (middle) in control and morphant explants. Overlays (right) show costaining of phalloidin. Bar, 20 µm. (m) Summary data (94–422 cells from two to three knockdowns) quantifying the number and size of labeled structures. (n) Lifetime analysis of focal adhesion kinase–GFP in control and morphant explants (30–58 focal adhesions from two knockdowns). 10 ng AMO1 was used for all knockdowns. Error bars represent SEM. *, P

    Journal: The Journal of Cell Biology

    Article Title: Ca2+/H+ exchange by acidic organelles regulates cell migration in vivo

    doi: 10.1083/jcb.201510019

    Figure Lengend Snippet: CAX regulates cell substrate attachment. (a and b) Effect of CAX knockdown on chemotaxis. (a) Tracks of individual explants migrating toward beads soaked in Sdf-1 over the entire period of recording (left) or the initial 100 min (right). (b) Summary data (from three knockdowns) quantifying forward motion index, persistence, and velocity of migration. (c–g) Effect of CAX knockdown on cell dispersion. (c) Summary data from three knockdowns quantifying the size of control and morphant explants 30 min after plating. (d) Representative color-coded triangulation diagrams at time 0 and 6 h from control or morphant explants. (e) Time courses (from three knockdowns) quantifying triangle area. Data are normalized to control explants at time 0. (f) Transmitted light micrographs showing spreading of control or morphant explants over a 20-min period. Panels on the right show an overlay of the area occupied by the explants where red represents the extent of spreading. Bar, 500 µm. (g) Summary data from three knockdowns quantifying spread area. (h and i) Effect of Ca 2+ buffering on cell dispersion. Triangulation (h) and spreading (i) analysis of explants treated with cell-permeable Ca 2+ chelators (50 µM BAPTA/EGTA-AM) or DMSO 30 min before imaging. (j and k) Live cell imaging of CAX. Stills from time-lapse confocal imaging of explants coexpressing mRFP-CAX and either membrane GFP (j; bar, 10 µm) or focal adhesion kinase–GFP (k). Arrowheads mark small CAX-positive vesicles. (k) Bars: (top) 4 µm; (bottom) 1 µm. Heat map summarizes the mean projection for CAX-positive vesicles relative to the centroid of focal adhesions (intersection of white lines) within 5 × 5–µm regions of interest. Data are from 28 focal adhesions from two expression experiments. (l–n) Effect of CAX knockdown on focal adhesions. (l) Expression of focal adhesion kinase–GFP (left) and immunocytochemistry analysis using an antibody to phosphopaxillin (middle) in control and morphant explants. Overlays (right) show costaining of phalloidin. Bar, 20 µm. (m) Summary data (94–422 cells from two to three knockdowns) quantifying the number and size of labeled structures. (n) Lifetime analysis of focal adhesion kinase–GFP in control and morphant explants (30–58 focal adhesions from two knockdowns). 10 ng AMO1 was used for all knockdowns. Error bars represent SEM. *, P

    Article Snippet: To test the effects of Ca2+ buffering, explants were incubated for 30 min with 50 µM BAPTA-AM (Cambridge Bioscience) or EGTA-AM (AnaSpec).

    Techniques: Chemotaxis Assay, Migration, Imaging, Live Cell Imaging, Expressing, Immunocytochemistry, 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

    Protoplast shrinkage at 55°C HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM EGTA prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p

    Journal: bioRxiv

    Article Title: Apoptosis in plants: from semantic appeal to empirical rejection

    doi: 10.1101/2020.09.26.314583

    Figure Lengend Snippet: Protoplast shrinkage at 55°C HS is an ATP- and Ca 2+ -independent process. A Mitochondria in BY-2 cells stained with MitoTracker Red and imaged 10 min after 55°C or 85°C HS, or after treatment with 48 µM CCCP. Severely damaged mitochondria were observed upon all three treatments. B Loss of intracellular ATP content upon HS. Snap freeze-thaw treatment in liquid nitrogen (N 2 ) and CCCP treatment were used as positive controls for completely disrupted and uncoupled mitochondria, respectively. The experiment was repeated twice, each time using four biological replicates per treatment. C MitoTracker Red staining of BY-2 cells exposed to 55°C in the presence or absence of 15 µM Cyclosporin A (CsA) reveals that inhibition of MPTP opening does not rescue mitochondria from severe damage and loss of membrane potential caused by HS. D MitoTracker Red localization in the cells pre-treated with 10 mM EGTA prior to the HS reveals that chelation of extracellular Ca 2+ does not rescue mitochondrial phenotype. E, F Dynamicsof cell death (% SO-positive cells; E ) and protoplast shrinkage (F) in cells with normal and uncoupled (48 µM CCCP treatment) mitochondria. G, H Pre-treatment with 10 mM EGTA before HS does not affect dynamics of cell death (% SO-positive cells; G ) and protoplast shrinkage (H) . Experiments shown in E-H were repeated three times, with ≥ 184 cells per treatment and time point. Each microscopy experiment was performed at least twice. Scale bars, 20 µm ( A ) or 50 µm ( C, D ). IQR, interquartile range. B, E - H , one-way ANOVA with Dunnet’s test; *, p

    Article Snippet: Four different treatments (all at room temperature) were applied prior to 10-min HS at 55°C: (i) 10 mM EGTA (Merck, E3889), pH 8.0 for 10 min, (ii) 0.1% DMSO for 2 h, (iii) 15 µM cyclosporin A (CsA) for 2 h, and (iv) 10 mM EGTA (applied 10 min prior to HS) and 15 µM CsA (applied 2 h prior to HS).

    Techniques: Staining, Inhibition, Microscopy

    HS-induced cell death is ATP- and Ca 2+ -independent process. A-D Morphology of FDA-stained cells under normal conditions (no HS) and after a 55°C HS. Protoplast shrinkage is denoted by arrows. Pre-treatment with 48 µM CCCP for 10 min (B) , 15 µM CsA for 2 h (C ), or 10 mM EGTA for 10 min ( D ) did not alleviate protoplast shrinkage upon HS. Each treatment was repeated at least twice. E, G Sytox Orange (SO) staining of BY-2 cells heat-shocked for 10 min at 40, 45, or 50°C and imaged after 6 h ( E ) and 24 h ( G ). Pre-treatment with CCCP provided no protection against cell death and protoplast shrinkage at any of the tested HS temperatures. F, H Quantification of cell death (% SO-positive cells) in the samples shown in E and G , respectively. DIC, differential interference contrast microscopy. IQR, interquartile range. Experiments shown in F and H were repeated three times, with ≥ 170 cells per treatment and time point. The data was subjected to one-way ANOVA with Bonferroni correction. Scale bars, 20 µm (A-D) or 100 µm (E, G) .

    Journal: bioRxiv

    Article Title: Apoptosis in plants: from semantic appeal to empirical rejection

    doi: 10.1101/2020.09.26.314583

    Figure Lengend Snippet: HS-induced cell death is ATP- and Ca 2+ -independent process. A-D Morphology of FDA-stained cells under normal conditions (no HS) and after a 55°C HS. Protoplast shrinkage is denoted by arrows. Pre-treatment with 48 µM CCCP for 10 min (B) , 15 µM CsA for 2 h (C ), or 10 mM EGTA for 10 min ( D ) did not alleviate protoplast shrinkage upon HS. Each treatment was repeated at least twice. E, G Sytox Orange (SO) staining of BY-2 cells heat-shocked for 10 min at 40, 45, or 50°C and imaged after 6 h ( E ) and 24 h ( G ). Pre-treatment with CCCP provided no protection against cell death and protoplast shrinkage at any of the tested HS temperatures. F, H Quantification of cell death (% SO-positive cells) in the samples shown in E and G , respectively. DIC, differential interference contrast microscopy. IQR, interquartile range. Experiments shown in F and H were repeated three times, with ≥ 170 cells per treatment and time point. The data was subjected to one-way ANOVA with Bonferroni correction. Scale bars, 20 µm (A-D) or 100 µm (E, G) .

    Article Snippet: Four different treatments (all at room temperature) were applied prior to 10-min HS at 55°C: (i) 10 mM EGTA (Merck, E3889), pH 8.0 for 10 min, (ii) 0.1% DMSO for 2 h, (iii) 15 µM cyclosporin A (CsA) for 2 h, and (iv) 10 mM EGTA (applied 10 min prior to HS) and 15 µM CsA (applied 2 h prior to HS).

    Techniques: Staining, Microscopy

    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

    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