Structured Review

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A. hydrophila infection leads to increased CaM expression in HKM. (A) HKM transfected separately with CaM-siRNA, scrambled siRNA or pre-treated with CMZ, Vp, Nf, <t>EGTA,</t> <t>BAPTA/AM</t> for different time periods were infected with A. hydrophila and CaM protein content measured in the lysates 2 h p.i. using EIA kit. HKM pre-treated with CMZ or transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and checked for (B) Hoechst 33342 positive cells and caspase-3 activity and (C) AV-PI staining 24 h p.i. (D) HKM transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and CaM mRNA expression detected by real time PCR 2 h p.i. Vertical bars represent mean ± SE (n = 6). * P
Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Role of Calmodulin-Calmodulin Kinase II, cAMP/Protein Kinase A and ERK 1/2 on Aeromonas hydrophila-Induced Apoptosis of Head Kidney Macrophages"

Article Title: Role of Calmodulin-Calmodulin Kinase II, cAMP/Protein Kinase A and ERK 1/2 on Aeromonas hydrophila-Induced Apoptosis of Head Kidney Macrophages

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1004018

A. hydrophila infection leads to increased CaM expression in HKM. (A) HKM transfected separately with CaM-siRNA, scrambled siRNA or pre-treated with CMZ, Vp, Nf, EGTA, BAPTA/AM for different time periods were infected with A. hydrophila and CaM protein content measured in the lysates 2 h p.i. using EIA kit. HKM pre-treated with CMZ or transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and checked for (B) Hoechst 33342 positive cells and caspase-3 activity and (C) AV-PI staining 24 h p.i. (D) HKM transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and CaM mRNA expression detected by real time PCR 2 h p.i. Vertical bars represent mean ± SE (n = 6). * P
Figure Legend Snippet: A. hydrophila infection leads to increased CaM expression in HKM. (A) HKM transfected separately with CaM-siRNA, scrambled siRNA or pre-treated with CMZ, Vp, Nf, EGTA, BAPTA/AM for different time periods were infected with A. hydrophila and CaM protein content measured in the lysates 2 h p.i. using EIA kit. HKM pre-treated with CMZ or transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and checked for (B) Hoechst 33342 positive cells and caspase-3 activity and (C) AV-PI staining 24 h p.i. (D) HKM transfected with CaM-siRNA or scrambled siRNA were infected with A. hydrophila and CaM mRNA expression detected by real time PCR 2 h p.i. Vertical bars represent mean ± SE (n = 6). * P

Techniques Used: Infection, Chick Chorioallantoic Membrane Assay, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Activity Assay, Staining, Real-time Polymerase Chain Reaction

2) Product Images from "Herpes simplex virus triggers activation of calcium-signaling pathways"

Article Title: Herpes simplex virus triggers activation of calcium-signaling pathways

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200301084

2-APB and BAPTA-AM prevent viral-induced FAK phosphorylation. CaSki cells were serum starved overnight, and a synchronized infection with HSV-2(G) was conducted as described in Materials and methods. 100 μM 2-APB, 10 μM nifedipine, or control buffer (5% DMSO) was added to cells at the time of binding or penetration (temperature shift, 15 min, 37°C). Cells were treated with low pH citrate buffer, washed, and cell lysates were prepared 10 min after citrate treatment. Western blots were prepared, probed with anti-FAK pY 397 , and scanned by optical densitometry. Blots were then stripped and reprobed with mAb to total FAK. A representative blot is shown in A. Alternatively, cells were pretreated for 2 h with BAPTA-AM, EGTA, or 5% methanol in PBS (control), and then exposed to HSV-2(G). (B) Lysates were prepared 5 min after infection and analyzed by Western blotting for FAK phosphorylation. (C) Results obtained from three independent experiments are summarized graphically as odu of phosphorylated FAK:odu total FAK as a percentage of control (HSV-2(G) in the absence of any inhibitors). Results are the mean ± SD obtained from three independent experiments; asterisks indicate P
Figure Legend Snippet: 2-APB and BAPTA-AM prevent viral-induced FAK phosphorylation. CaSki cells were serum starved overnight, and a synchronized infection with HSV-2(G) was conducted as described in Materials and methods. 100 μM 2-APB, 10 μM nifedipine, or control buffer (5% DMSO) was added to cells at the time of binding or penetration (temperature shift, 15 min, 37°C). Cells were treated with low pH citrate buffer, washed, and cell lysates were prepared 10 min after citrate treatment. Western blots were prepared, probed with anti-FAK pY 397 , and scanned by optical densitometry. Blots were then stripped and reprobed with mAb to total FAK. A representative blot is shown in A. Alternatively, cells were pretreated for 2 h with BAPTA-AM, EGTA, or 5% methanol in PBS (control), and then exposed to HSV-2(G). (B) Lysates were prepared 5 min after infection and analyzed by Western blotting for FAK phosphorylation. (C) Results obtained from three independent experiments are summarized graphically as odu of phosphorylated FAK:odu total FAK as a percentage of control (HSV-2(G) in the absence of any inhibitors). Results are the mean ± SD obtained from three independent experiments; asterisks indicate P

Techniques Used: Infection, Binding Assay, Western Blot

The ER Ca 2+ release is required for HSV infection. The effects of pretreating cells for 1 h with 50 μM BAPTA-AM, 0.5 mM EGTA, or adding 100 μM 2-APB, 10 μM nifedipine, or 10 μM verapamil during viral penetration on HSV-1(KOS), HSV-2(G), or VSV infection of Vero cells (top) or CaSki cells (bottom) were examined. Results are presented as pfu formed in the presence of the indicated concentration of drug as a percentage of pfu formed in cells treated with control buffer (5% DMSO for pharmacological inhibitors or 5% methanol for BAPTA), and are means of at least three independent experiments conducted in duplicate. Cytotoxicity of pharmacological agents was determined in parallel after a 2-h exposure to drug and quantifying viable, proliferating cells at 24 h by MTS assay. Cell viability results are expressed as odu reading in the presence of the drug as a percentage of odu reading in the presence of control buffer, and are means of two independent experiments conducted in triplicate. Asterisks denote P
Figure Legend Snippet: The ER Ca 2+ release is required for HSV infection. The effects of pretreating cells for 1 h with 50 μM BAPTA-AM, 0.5 mM EGTA, or adding 100 μM 2-APB, 10 μM nifedipine, or 10 μM verapamil during viral penetration on HSV-1(KOS), HSV-2(G), or VSV infection of Vero cells (top) or CaSki cells (bottom) were examined. Results are presented as pfu formed in the presence of the indicated concentration of drug as a percentage of pfu formed in cells treated with control buffer (5% DMSO for pharmacological inhibitors or 5% methanol for BAPTA), and are means of at least three independent experiments conducted in duplicate. Cytotoxicity of pharmacological agents was determined in parallel after a 2-h exposure to drug and quantifying viable, proliferating cells at 24 h by MTS assay. Cell viability results are expressed as odu reading in the presence of the drug as a percentage of odu reading in the presence of control buffer, and are means of two independent experiments conducted in triplicate. Asterisks denote P

Techniques Used: Infection, Concentration Assay, MTS Assay

3) Product Images from "Areca Nut Extract Induces Pyknotic Necrosis in Serum-Starved Oral Cells via Increasing Reactive Oxygen Species and Inhibiting GSK3?: An Implication for Cytopathic Effects in Betel Quid Chewers"

Article Title: Areca Nut Extract Induces Pyknotic Necrosis in Serum-Starved Oral Cells via Increasing Reactive Oxygen Species and Inhibiting GSK3?: An Implication for Cytopathic Effects in Betel Quid Chewers

Journal: PLoS ONE

doi: 10.1371/journal.pone.0063295

Calcium signaling was involved in ANE-induced pyknotic necrosis. (A) Calcium flux in SAS cells treated with the indicated doses of ANE was measured as described in experimental procedures. (B) Similar to the condition in (A), calcium flux was evaluated in the presence or absence of NAC. (C) Cells co-treated with ANE and either 5 mM EGTA or 5 µM BAPTA-AM were photographed at the indicated time points.
Figure Legend Snippet: Calcium signaling was involved in ANE-induced pyknotic necrosis. (A) Calcium flux in SAS cells treated with the indicated doses of ANE was measured as described in experimental procedures. (B) Similar to the condition in (A), calcium flux was evaluated in the presence or absence of NAC. (C) Cells co-treated with ANE and either 5 mM EGTA or 5 µM BAPTA-AM were photographed at the indicated time points.

Techniques Used:

Related Articles

other:

Article Title: Spontaneous miniature outward currents in mechanically dissociated rat Meynert neurons
Article Snippet: However, the SMOC amplitude was never increased by applying EGTA-AM or BAPTA-AM ( ).

Article Title: HSP70 Acetylation Prevents Combined mTORC1/2 Inhibitor and Curcumin Treatment-Induced Apoptosis
Article Snippet: Calbiochem supplied EGTA-AM (San Diego, CA, USA).

Article Title: Myosin V functions as a vesicle tether at the plasma membrane to control neurotransmitter release in central synapses
Article Snippet: Pharmacology MyoVin-1 (Millipore), Pentabromopseudalin (PBP, Fisher Scientific) or EGTA-AM (Millipore) were diluted in DMSO (Sigma-Aldrich) and stored at −20°C.

Article Title: Activity-Dependent Maintenance of Long-Term Potentiation at Visual Cortical Inhibitory Synapses
Article Snippet: The compounds used were obtained from the following sources: APV, DNQX, and from Tocris Cookson (Bristol, UK); iberiotoxin, tetrodotoxin (TTX), bicuculline methiodide, noradrenaline, and serotonin from Sigma (St. Louis, MO); 4-aminopyridine, charybdotoxin, ω-conotoxin GVIA, and nifedipine from Research Biochemicals (Natick, MA); ω-agatoxin IVA from Peptide Institute (Osaka, Japan); EGTA-AM from Calbiochem (La Jolla, CA); saxitoxin (STX) from Alexis (San Diego, CA); and isoflurane from Abbott Laboratories (North Chicago, IL).

Article Title: Salt-Induced Expression of the Vacuolar H+-ATPase in the Common Ice Plant Is Developmentally Controlled and Tissue Specific 1
Article Snippet: The solution was supplemented with 400 m m NaCl or 400 m m NaCl and effectors, i.e. 20 m m EGTA/800 μ m EGTA/AM (Calbiochem, La Jolla, CA), 50 μ m neomycin sulfate (Calbiochem), 400 μ m forskolin (Calbiochem), 10 μ m mastoparan (Sigma, St. Louis), or 120 n m cholera toxin (Calbiochem).

Article Title: Calbindin-D28K Limits Dopamine Release in Ventral but Not Dorsal Striatum by Regulating Ca2+ Availability and Dopamine Transporter Function
Article Snippet: Drugs and Solutions BAPTA-AM and DHβE were purchased from Ascent Scientific or Tocris UK; pluronic acid was from Life Technologies; EGTA-AM was from Millipore.

Inhibition:

Article Title: Characterizing the Site and Mode of Action of Dynorphin at Hippocampal Mossy Fiber Synapses in the Guinea Pig
Article Snippet: .. Although EGTA-AM had no effect on the inhibitory action of dynorphin, heterosynaptic inhibition was severely depressed in these slices, demonstrating that the release of dynorphin requires a rise in Ca2+ in the mossy fiber terminal. .. The long duration of heterosynaptic inhibition could result from a number of factors, such as prolonged release of dynorphin, prolonged presence of the peptide in the extracellular space, or prolonged second messenger activity.

Activation Assay:

Article Title: Activation of AMPK by Bitter Melon Triterpenoids Involves CaMKK?
Article Snippet: Accordingly, incubation of L6 myotubes with BMT-17 significantly increased the AMPK activity. .. Pre-incubation with EGTA-AM to deplete the intracellular calcium levels did not alter the BMT-17-induced AMPK activation ( ). ..

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    Millipore egta
    Ca 2+ chelators and Ca 2+ channel blockers inhibit KSHV-induced Ang-2 release. Identical numbers of <t>HUVECs</t> were pretreated with the Ca 2+ chelators BAPTA (100 μM), BAPTA-AM (100 μM), and <t>EGTA</t> (100 μM); the Ca 2+ channel blockers nifedipine
    Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ca 2+ chelators and Ca 2+ channel blockers inhibit KSHV-induced Ang-2 release. Identical numbers of HUVECs were pretreated with the Ca 2+ chelators BAPTA (100 μM), BAPTA-AM (100 μM), and EGTA (100 μM); the Ca 2+ channel blockers nifedipine

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Induces Rapid Release of Angiopoietin-2 from Endothelial Cells

    doi: 10.1128/JVI.03303-12

    Figure Lengend Snippet: Ca 2+ chelators and Ca 2+ channel blockers inhibit KSHV-induced Ang-2 release. Identical numbers of HUVECs were pretreated with the Ca 2+ chelators BAPTA (100 μM), BAPTA-AM (100 μM), and EGTA (100 μM); the Ca 2+ channel blockers nifedipine

    Article Snippet: To test if protein tyrosine phosphorylation is also involved in rapid Ang-2 release by other known factors, such as thrombin , we pretreated HUVECs with 2 μM genistein for 2 h, followed by stimulation with 2 U/ml thrombin purified from human plasma (Sigma-Aldrich) for 15 min. To examine the effects of Ca2+ mobilization on Ang-2 release, HUVECs were pretreated with 100 μM Ca2+ chelator BAPTA, BAPTA-AM (Sigma-Aldrich), or EGTA (Sigma-Aldrich); 20 μM Ca2+ entry blocker SKF 96365 (abcamBiochemicals, Cambrige, MA); 5 μM Ca2+ channel blocker nifedipine (Sigma-Aldrich) or amlodipine (Pfizer, New York, NY); and 5 μM thapsigargin (Sigma-Aldrich) for 30 min, followed by exposure to viral stock solution for 15 min.

    Techniques:

    (A) Polymerization of 1.5 μM 100% pyrene labeled skeletal Mg-ATP-G-actin (5 mM Tris, pH 8, 0.1 mM MgCl 2 , 0.2 mM EGTA, 0.2 mM ATP, 0.5 mM DTT). Additions: 1 ) 4.0 μM polylysine containing 0.2 mM MgCl 2 ; 2 ) 4.0 μM lysozyme; 3 ) 0.5 mM spermine; 4 ) 2.0 mM MgCl 2 . The polymerization was followed in a PTI spectrofluorometer (see Materials and Methods). (B) Actin polymerization of 2.0 μM pyrene labeled Mg-ATP-G-actin by 4.0 μM polylysine and 0.2 mM MgCl 2 at pH 8 (G buffer) followed in a stopped-flow fluorometer (see Materials and Methods). (C) Excimer formation during the polymerization of 2.0 μM pyrene labeled (100%) Mg-ATP-G- actin by 4 μM polylysine and 0.15 mM MgCl 2 at pH 8.0 followed in a stopped-flow fluorometer (see Materials and Methods). (D) Kinetics of actin filaments elongation (green) and excimer fluorescence decay (red). 4.0 μM polylysine and 0.1 mM MgCl 2 were added to 1.5 μM 100% pyrene labeled MgATP-G-actin. Excimer fluorescence decay started 9 s after polylysine addition but was shifted to 0 s on this figure. The elongation and excimer decay data were fitted to a single and two exponential equations (black lines) with rate constants 0.054 s− 1 for elongation and 0.037 and 0.0005 s− 1 for excimer decay. Fluorescence change is shown as a % of total change upon elongation (365 nm excitation and 386 nm emission) and excimer formation (343 nm excitation and 478 nm emission wavelengths).

    Journal: Biochemistry

    Article Title: Antiparallel dimer and actin assembly

    doi: 10.1021/bi1002663

    Figure Lengend Snippet: (A) Polymerization of 1.5 μM 100% pyrene labeled skeletal Mg-ATP-G-actin (5 mM Tris, pH 8, 0.1 mM MgCl 2 , 0.2 mM EGTA, 0.2 mM ATP, 0.5 mM DTT). Additions: 1 ) 4.0 μM polylysine containing 0.2 mM MgCl 2 ; 2 ) 4.0 μM lysozyme; 3 ) 0.5 mM spermine; 4 ) 2.0 mM MgCl 2 . The polymerization was followed in a PTI spectrofluorometer (see Materials and Methods). (B) Actin polymerization of 2.0 μM pyrene labeled Mg-ATP-G-actin by 4.0 μM polylysine and 0.2 mM MgCl 2 at pH 8 (G buffer) followed in a stopped-flow fluorometer (see Materials and Methods). (C) Excimer formation during the polymerization of 2.0 μM pyrene labeled (100%) Mg-ATP-G- actin by 4 μM polylysine and 0.15 mM MgCl 2 at pH 8.0 followed in a stopped-flow fluorometer (see Materials and Methods). (D) Kinetics of actin filaments elongation (green) and excimer fluorescence decay (red). 4.0 μM polylysine and 0.1 mM MgCl 2 were added to 1.5 μM 100% pyrene labeled MgATP-G-actin. Excimer fluorescence decay started 9 s after polylysine addition but was shifted to 0 s on this figure. The elongation and excimer decay data were fitted to a single and two exponential equations (black lines) with rate constants 0.054 s− 1 for elongation and 0.037 and 0.0005 s− 1 for excimer decay. Fluorescence change is shown as a % of total change upon elongation (365 nm excitation and 386 nm emission) and excimer formation (343 nm excitation and 478 nm emission wavelengths).

    Article Snippet: ATP, poly-L-lysine (MW 4000), dithiotreitol (DTT), N-ethylmaleimide (NEM), spermine and EGTA were purchased from Sigma Chemical Co. (St Louis, MO).

    Techniques: Labeling, Flow Cytometry, Fluorescence

    Activity‐dependent bulk endocytosis ( ADBE ) is abolished by both BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min before loading with tetramethylrhodamine‐dextran ( TMR )‐dextran (50 μM) during stimulation with a train of 800 action potentials (80 Hz). (a) Quantification of the number of evoked TMR ‐dextran puncta per field of view in cultures treated with either DMSO (Ctrl, blue bars) BAPTA ‐ AM (red bars) or EGTA ‐ AM (purple bars) ± SEM . *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: Activity‐dependent bulk endocytosis ( ADBE ) is abolished by both BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min before loading with tetramethylrhodamine‐dextran ( TMR )‐dextran (50 μM) during stimulation with a train of 800 action potentials (80 Hz). (a) Quantification of the number of evoked TMR ‐dextran puncta per field of view in cultures treated with either DMSO (Ctrl, blue bars) BAPTA ‐ AM (red bars) or EGTA ‐ AM (purple bars) ± SEM . *** p

    Article Snippet: This dephosphorylation is arrested in cultures treated with either BAPTA‐AM or EGTA‐AM (Fig. ).

    Techniques: Activity Assay

    Dynamin I dephosphorylation is arrested by BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min. They were then either stimulated with a train of 800 action potentials (80 Hz) or left to rest for 10 s. Cultures were then immediately lysed in SDS sample buffer. (a) Representative immunoblots of cerebellar granule neuron lysates probed with either dynamin I phospho‐Ser774 ( PD ynI) or synaptophysin (Syp) antibodies. (b) Quantification of PD ynI levels after normalisation against Syp ( PD ynI/Syp) ± SEM . Blue bars represent Ctrl, red bars BAPTA ‐ AM and purple bars EGTA ‐ AM , *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: Dynamin I dephosphorylation is arrested by BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min. They were then either stimulated with a train of 800 action potentials (80 Hz) or left to rest for 10 s. Cultures were then immediately lysed in SDS sample buffer. (a) Representative immunoblots of cerebellar granule neuron lysates probed with either dynamin I phospho‐Ser774 ( PD ynI) or synaptophysin (Syp) antibodies. (b) Quantification of PD ynI levels after normalisation against Syp ( PD ynI/Syp) ± SEM . Blue bars represent Ctrl, red bars BAPTA ‐ AM and purple bars EGTA ‐ AM , *** p

    Article Snippet: This dephosphorylation is arrested in cultures treated with either BAPTA‐AM or EGTA‐AM (Fig. ).

    Techniques: De-Phosphorylation Assay, Western Blot

    SV exocytosis is arrested by BAPTA ‐ AM but not by EGTA ‐ AM . (a) Schematic of the experimental design. Cultures were loaded with 10 μM FM 1‐43 during a train of 800 action potentials (80 Hz). After a 10 min rest period FM 1‐43 was unloaded with a second 800 action potential stimulus (S1). After a 10 min rest period, the same loading/unloading protocol was performed again, with the exception being that cultures were incubated with either BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or dimethylsulfoxide ( DMSO ) (Ctrl) for 30 min directly after S2 loading. (b) Representative traces showing the S2 unloading profiles of Ctrl (blue circles), BAPTA ‐ AM (red circles) and EGTA ‐ AM (purple circles) ‐treated cultures. (c) Quantification of the extent of FM 1‐43 unloading at both S1 (ΔS1) and S2 (ΔS2) presented as the ratio ΔS2/ΔS1 ± SEM . *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: SV exocytosis is arrested by BAPTA ‐ AM but not by EGTA ‐ AM . (a) Schematic of the experimental design. Cultures were loaded with 10 μM FM 1‐43 during a train of 800 action potentials (80 Hz). After a 10 min rest period FM 1‐43 was unloaded with a second 800 action potential stimulus (S1). After a 10 min rest period, the same loading/unloading protocol was performed again, with the exception being that cultures were incubated with either BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or dimethylsulfoxide ( DMSO ) (Ctrl) for 30 min directly after S2 loading. (b) Representative traces showing the S2 unloading profiles of Ctrl (blue circles), BAPTA ‐ AM (red circles) and EGTA ‐ AM (purple circles) ‐treated cultures. (c) Quantification of the extent of FM 1‐43 unloading at both S1 (ΔS1) and S2 (ΔS2) presented as the ratio ΔS2/ΔS1 ± SEM . *** p

    Article Snippet: This dephosphorylation is arrested in cultures treated with either BAPTA‐AM or EGTA‐AM (Fig. ).

    Techniques: Incubation

    ( A – C ) Maximal projection of image z-stack of MDCK cells treated with blebbistatin on ( A ) a flat substrate, a cylinder substrate of radius of curvature R c , ( B ) 40 μ m, and ( C ) 20 μ m, with corresponding xz-cross-sectional view below each image. Blue corresponds to nucleus, green corresponds to E-cadherin, and red corresponds to F-actin. Scale bar represents 20 μ m. ( D ) Angular distribution of MDCK cells treated with blebbistatin on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( E – G ) Maximal projection of image z-stack of MDCK cells treated with calyculin A on ( E ) a flat substrate, a cylinder substrate of radius of curvature R c , ( F ) 40 μ m, and ( G ) 20 μ m, with corresponding xz-cross-sectional view below each image, is shown. Scale bar represents 20 μ m. ( H ) Angular distribution of MDCK cells treated with calyculin A, on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( I – K ) Maximal projection of image z-stack of MDCK cells treated with EGTA on ( I ) a flat substrate, a cylinder substrate of radius of curvature R c , ( J ) 40 μ m, and ( K ) 20 μ m, with corresponding xz-cross-sectional view below each image, is shown. Scale bar represents 20 μ m. ( L ) Angular distribution of MDCK cells treated with EGTA, on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( M ) Percentage of MDCK cells with various treatment that lie within 20° of the axis of the channel versus radius of curvature R c is shown. Error bars indicate standard error of the mean. ( N ) Elongation of MDCK cells with various treatment versus radius of curvature R c is shown . ∗ and ∗∗∗ represent p

    Journal: Biophysical Journal

    Article Title: Cell-Cell Adhesion and Cortical Actin Bending Govern Cell Elongation on Negatively Curved Substrates

    doi: 10.1016/j.bpj.2018.02.027

    Figure Lengend Snippet: ( A – C ) Maximal projection of image z-stack of MDCK cells treated with blebbistatin on ( A ) a flat substrate, a cylinder substrate of radius of curvature R c , ( B ) 40 μ m, and ( C ) 20 μ m, with corresponding xz-cross-sectional view below each image. Blue corresponds to nucleus, green corresponds to E-cadherin, and red corresponds to F-actin. Scale bar represents 20 μ m. ( D ) Angular distribution of MDCK cells treated with blebbistatin on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( E – G ) Maximal projection of image z-stack of MDCK cells treated with calyculin A on ( E ) a flat substrate, a cylinder substrate of radius of curvature R c , ( F ) 40 μ m, and ( G ) 20 μ m, with corresponding xz-cross-sectional view below each image, is shown. Scale bar represents 20 μ m. ( H ) Angular distribution of MDCK cells treated with calyculin A, on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( I – K ) Maximal projection of image z-stack of MDCK cells treated with EGTA on ( I ) a flat substrate, a cylinder substrate of radius of curvature R c , ( J ) 40 μ m, and ( K ) 20 μ m, with corresponding xz-cross-sectional view below each image, is shown. Scale bar represents 20 μ m. ( L ) Angular distribution of MDCK cells treated with EGTA, on substrates of various radius of curvature R c , with 0° and 90° being perpendicular and parallel to the long axis of the cylinder, is shown. ( M ) Percentage of MDCK cells with various treatment that lie within 20° of the axis of the channel versus radius of curvature R c is shown. Error bars indicate standard error of the mean. ( N ) Elongation of MDCK cells with various treatment versus radius of curvature R c is shown . ∗ and ∗∗∗ represent p

    Article Snippet: For cells treated with EGTA, blebbistatin, or calyculin A, final concentrations of 4 mM EGTA (Sigma), 100 μL/mL blebbistatin (Tocris Bioscience, Bristol, United Kingdom), or 10 nM calyculin A (Sigma) were added to the respective samples 4 h before fixing.

    Techniques: