Structured Review

Millipore amiloride
Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of <t>amiloride</t> (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.
Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 105 article reviews
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amiloride - by Bioz Stars, 2020-02
99/100 stars

Images

1) Product Images from "Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice"

Article Title: Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0057317

Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of amiloride (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.
Figure Legend Snippet: Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of amiloride (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.

Techniques Used: Mouse Assay, Concentration Assay

Cutoff point determination for nasal potential difference parameters distinguishing between FVB WT and F508del-CFTR mice. Receiver operating characteristics (ROC) curves for (A) baseline potential difference (V TE ), (B) amiloride response (ΔV TEAmil. ) and (C) low Cl - solution response (ΔV TElowCl - ), for WT and F508del-CFTR FVB mice are shown. AUC: area under the curve; 95% CI: 95% confidence interval. Cutoff points were determined by the best positive likelihood ratio of sensitivity/(1-specificity).
Figure Legend Snippet: Cutoff point determination for nasal potential difference parameters distinguishing between FVB WT and F508del-CFTR mice. Receiver operating characteristics (ROC) curves for (A) baseline potential difference (V TE ), (B) amiloride response (ΔV TEAmil. ) and (C) low Cl - solution response (ΔV TElowCl - ), for WT and F508del-CFTR FVB mice are shown. AUC: area under the curve; 95% CI: 95% confidence interval. Cutoff points were determined by the best positive likelihood ratio of sensitivity/(1-specificity).

Techniques Used: Mouse Assay

2) Product Images from "Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways"

Article Title: Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways

Journal: Hearing research

doi: 10.1016/j.heares.2009.03.001

The mechanosensory channel blocker amiloride prevents hair cell death induced by either acute or chronic treatment with either neomycin or gentamicin. A, Zebrafish larvae were pre-treated 1 mM amiloride then exposed concurrently to 200 μM neomycin or gentamicin acutely (30 min aminoglycoside exposure followed by 1 hour washout). Controls were treated with aminoglycoside without amiloride exposure. There is a significant reduction in hair cell death in the presence of amiloride for both aminoglycosides (2-way ANOVA, p
Figure Legend Snippet: The mechanosensory channel blocker amiloride prevents hair cell death induced by either acute or chronic treatment with either neomycin or gentamicin. A, Zebrafish larvae were pre-treated 1 mM amiloride then exposed concurrently to 200 μM neomycin or gentamicin acutely (30 min aminoglycoside exposure followed by 1 hour washout). Controls were treated with aminoglycoside without amiloride exposure. There is a significant reduction in hair cell death in the presence of amiloride for both aminoglycosides (2-way ANOVA, p

Techniques Used:

3) Product Images from "mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity"

Article Title: mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI73935

Inhibition of ENaC with amiloride blunts the natriuretic effect of PP242.
Figure Legend Snippet: Inhibition of ENaC with amiloride blunts the natriuretic effect of PP242.

Techniques Used: Inhibition

4) Product Images from "Insulin ameliorates pulmonary edema through the upregulation of epithelial sodium channel via the PI3K/SGK1 pathway in mice with lipopolysaccharide-induced lung injury"

Article Title: Insulin ameliorates pulmonary edema through the upregulation of epithelial sodium channel via the PI3K/SGK1 pathway in mice with lipopolysaccharide-induced lung injury

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2019.9809

Effect of exogenous insulin on pulmonary edema and alveolar fluid clearance in mice with LPS-induced lung injury (n=6 per group). (A) Wet-to-dry ratio and (B) alveolar fluid clearance at 0, 6, 12 and 24 h in the LPS-induced lung injury group. (C) Wet-to-dry ratio and (D) alveolar fluid clearance at 24 h in the LPS-induced lung injury group. Albumin solution containing amiloride (5×10 −4 M) was injected into the alveolar spaces 6 h after LPS administration. Data are presented as the mean ± standard error of the mean. Δ P
Figure Legend Snippet: Effect of exogenous insulin on pulmonary edema and alveolar fluid clearance in mice with LPS-induced lung injury (n=6 per group). (A) Wet-to-dry ratio and (B) alveolar fluid clearance at 0, 6, 12 and 24 h in the LPS-induced lung injury group. (C) Wet-to-dry ratio and (D) alveolar fluid clearance at 24 h in the LPS-induced lung injury group. Albumin solution containing amiloride (5×10 −4 M) was injected into the alveolar spaces 6 h after LPS administration. Data are presented as the mean ± standard error of the mean. Δ P

Techniques Used: Mouse Assay, Injection

5) Product Images from "Evidence for ammonium conductance in a mouse thick ascending limb cell line. Evidence for ammonium conductance in a mouse thick ascending limb cell line"

Article Title: Evidence for ammonium conductance in a mouse thick ascending limb cell line. Evidence for ammonium conductance in a mouse thick ascending limb cell line

Journal: Physiological Reports

doi: 10.14814/phy2.13379

NH 4 + conductance in oocytes injected with water or poly(A) RNA of ST ‐1 cells. (A and B) I – V relationships of NH 4 + currents measured by two‐electrode voltage clamp. Steady‐state currents were recorded before and after switching solutions from 20 mmol/L LiCl to the equimolar concentration of NH 4 Cl ( n = 7 RNA ‐injected oocytes and 5 water‐injected controls). (C) Mean slope of the I – V curve. Slope was determined near zero‐current voltage. (D) NH 4 + conductance ( G NH 4 ). G NH 4 represents the difference between the two slopes before and after NH 4 Cl application. (E) Effect of 10 μ mol/L amiloride on the NH 4 Cl‐induced slope. Data are presented as fold change relative to the slope before amiloride treatment ( n = 6 for each).
Figure Legend Snippet: NH 4 + conductance in oocytes injected with water or poly(A) RNA of ST ‐1 cells. (A and B) I – V relationships of NH 4 + currents measured by two‐electrode voltage clamp. Steady‐state currents were recorded before and after switching solutions from 20 mmol/L LiCl to the equimolar concentration of NH 4 Cl ( n = 7 RNA ‐injected oocytes and 5 water‐injected controls). (C) Mean slope of the I – V curve. Slope was determined near zero‐current voltage. (D) NH 4 + conductance ( G NH 4 ). G NH 4 represents the difference between the two slopes before and after NH 4 Cl application. (E) Effect of 10 μ mol/L amiloride on the NH 4 Cl‐induced slope. Data are presented as fold change relative to the slope before amiloride treatment ( n = 6 for each).

Techniques Used: Injection, Concentration Assay

NH 4 + currents in ST ‐1 cells measured by whole cell patch clamp. (A) An example of a whole cell current evoked by 20 mmol/L NH 4 Cl in an ST ‐1 cell. The recording was performed in the presence of 2 mmol/L BaCl 2 , 10 mmol/L TEA , 5 mmol/L BAPTA . The holding potential was −70 mV . NH 4 Cl replaced LiCl at the equimolar concentration. (B) Mean peak current. The difference in currents before and after NH 4 Cl application represents an NH 4 Cl‐mediated current ( n = 9). (C) I – V relationships. Peak or steady‐state currents at different voltages were acquired by the voltage command stepping from −80 to +15 mV ( n = 5). The difference between the two I – V curves is shown in a gray line. (D) Mean slope determined from the I – V curves. (E) Effect of 1 μ mol/L amiloride on NH 4 Cl‐induced currents ( I NH 4Cl ). Data are presented as fold change relative to I NH 4Cl produced without amiloride ( n = 6 for each).
Figure Legend Snippet: NH 4 + currents in ST ‐1 cells measured by whole cell patch clamp. (A) An example of a whole cell current evoked by 20 mmol/L NH 4 Cl in an ST ‐1 cell. The recording was performed in the presence of 2 mmol/L BaCl 2 , 10 mmol/L TEA , 5 mmol/L BAPTA . The holding potential was −70 mV . NH 4 Cl replaced LiCl at the equimolar concentration. (B) Mean peak current. The difference in currents before and after NH 4 Cl application represents an NH 4 Cl‐mediated current ( n = 9). (C) I – V relationships. Peak or steady‐state currents at different voltages were acquired by the voltage command stepping from −80 to +15 mV ( n = 5). The difference between the two I – V curves is shown in a gray line. (D) Mean slope determined from the I – V curves. (E) Effect of 1 μ mol/L amiloride on NH 4 Cl‐induced currents ( I NH 4Cl ). Data are presented as fold change relative to I NH 4Cl produced without amiloride ( n = 6 for each).

Techniques Used: Patch Clamp, Concentration Assay, Produced

Related Articles

Diagnostic Assay:

Article Title: mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity
Article Snippet: Urine samples were sent to Stanford University Department of Comparative Medicine Diagnostic Laboratory for determination of urine parameters. .. In the afternoon of day 3 of acclimatization, for studies with amiloride, mice were given 20 mg/l amiloride (Sigma-Aldrich) in their drinking water.

Flow Cytometry:

Article Title: Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice
Article Snippet: .. The following solutions were perfused through the second lumen at a flow rate of 1.5 ml/h: (1) Cl- solution for basal measurement, (2) 100 µM amiloride in Cl- solution (Sigma-Aldrich, USA), to block ENaC Na+ absorption (3) a low-Cl- solution (140 mM sodium gluconate, 6 mM potassium gluconate, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2 , 6 mM calcium gluconate, pH adjusted to 7.4 with NaOH) containing amiloride (100 µM) to drive Cl- secretion. ..

Patch Clamp:

Article Title: Evidence for ammonium conductance in a mouse thick ascending limb cell line. Evidence for ammonium conductance in a mouse thick ascending limb cell line
Article Snippet: Paragraph title: Whole cell patch clamp of ST‐1 cells ... For amiloride experiments, currents induced by NH4 Cl were recorded in the absence and then presence of 1 μ mol/L amiloride (Sigma‐Aldrich; Cat#: A7410).

Isolation:

Article Title: Insulin ameliorates pulmonary edema through the upregulation of epithelial sodium channel via the PI3K/SGK1 pathway in mice with lipopolysaccharide-induced lung injury
Article Snippet: Briefly, the isolated right lungs were placed in a humidified incubator at 37°C and ventilated with 100% nitrogen to remove oxygen from the alveolar spaces. .. Albumin solution containing amiloride (5×10−4 M; Sigma-Aldrich; Merck KGaA) was injected into the alveolar spaces.

Transferring:

Article Title: Evidence for ammonium conductance in a mouse thick ascending limb cell line. Evidence for ammonium conductance in a mouse thick ascending limb cell line
Article Snippet: The pipette had 116 KCl, 1.2 MgCl2 , 6 glucose, 10 HEPES, 10 TEA, 10 BAPTA, and 0.01 BaCl2 (pH 7.2). .. For amiloride experiments, currents induced by NH4 Cl were recorded in the absence and then presence of 1 μ mol/L amiloride (Sigma‐Aldrich; Cat#: A7410).

Blocking Assay:

Article Title: Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice
Article Snippet: .. The following solutions were perfused through the second lumen at a flow rate of 1.5 ml/h: (1) Cl- solution for basal measurement, (2) 100 µM amiloride in Cl- solution (Sigma-Aldrich, USA), to block ENaC Na+ absorption (3) a low-Cl- solution (140 mM sodium gluconate, 6 mM potassium gluconate, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2 , 6 mM calcium gluconate, pH adjusted to 7.4 with NaOH) containing amiloride (100 µM) to drive Cl- secretion. ..

Mouse Assay:

Article Title: mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity
Article Snippet: .. In the afternoon of day 3 of acclimatization, for studies with amiloride, mice were given 20 mg/l amiloride (Sigma-Aldrich) in their drinking water. .. The following morning, mice were injected i.p. with either 5 mg/kg amiloride in vehicle or 5 mg/kg amiloride plus 30 mg/kg PP242; the vehicle used was the same as described above.

Concentration Assay:

Article Title: The influence of mode of delivery, hormonal status and postnatal O2 environment on epithelial sodium channel (ENaC) expression in perinatal guinea-pig lung
Article Snippet: .. To determine the fractional inhibition of lung fluid clearance by amiloride, an excess of amiloride (10−3 M; Sigma) was added to the 5% albumin instillate solution to ensure an appropriate concentration at the alveolar epithelium, since a fraction is protein bound and another fraction rapidly leaves the airspace due to its low molecular weight. .. Clearance of fluid from the distal airspaces of the lungs was measured by the increase in protein concentration of the instilled solution over 1 h. Data are presented as a ratio between final and instilled protein concentration.

Sampling:

Article Title: The influence of mode of delivery, hormonal status and postnatal O2 environment on epithelial sodium channel (ENaC) expression in perinatal guinea-pig lung
Article Snippet: A sample of remaining lung fluid was aspirated by gently advancing a sampling catheter into a wedged position in a distal airway in the lungs. .. To determine the fractional inhibition of lung fluid clearance by amiloride, an excess of amiloride (10−3 M; Sigma) was added to the 5% albumin instillate solution to ensure an appropriate concentration at the alveolar epithelium, since a fraction is protein bound and another fraction rapidly leaves the airspace due to its low molecular weight.

Inhibition:

Article Title: The influence of mode of delivery, hormonal status and postnatal O2 environment on epithelial sodium channel (ENaC) expression in perinatal guinea-pig lung
Article Snippet: .. To determine the fractional inhibition of lung fluid clearance by amiloride, an excess of amiloride (10−3 M; Sigma) was added to the 5% albumin instillate solution to ensure an appropriate concentration at the alveolar epithelium, since a fraction is protein bound and another fraction rapidly leaves the airspace due to its low molecular weight. .. Clearance of fluid from the distal airspaces of the lungs was measured by the increase in protein concentration of the instilled solution over 1 h. Data are presented as a ratio between final and instilled protein concentration.

Labeling:

Article Title: Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways
Article Snippet: Hair cell survival was assessed with DASPEI labeling. .. For experiments with amiloride and acute aminoglycoside exposure, zebrafish larvae were pretreated for 15 minutes with 1 mM amiloride (Sigma) in embryo medium, transferred to 200 μM neomycin or gentamicin plus 1 mM amiloride for 30 min, washed four times in embryo medium and held in the final rinse for one hour prior to evaluation of hair cells with DASPEI.

Inverted Microscopy:

Article Title: Evidence for ammonium conductance in a mouse thick ascending limb cell line. Evidence for ammonium conductance in a mouse thick ascending limb cell line
Article Snippet: A coverslip on which cells were grown was mounted on a recording chamber affixed on the stage of a Nikon Eclipse T1 inverted microscope, equipped with a MP‐225 motorized micromanipulator. .. For amiloride experiments, currents induced by NH4 Cl were recorded in the absence and then presence of 1 μ mol/L amiloride (Sigma‐Aldrich; Cat#: A7410).

Protein Concentration:

Article Title: The influence of mode of delivery, hormonal status and postnatal O2 environment on epithelial sodium channel (ENaC) expression in perinatal guinea-pig lung
Article Snippet: To determine the fractional inhibition of lung fluid clearance by amiloride, an excess of amiloride (10−3 M; Sigma) was added to the 5% albumin instillate solution to ensure an appropriate concentration at the alveolar epithelium, since a fraction is protein bound and another fraction rapidly leaves the airspace due to its low molecular weight. .. Clearance of fluid from the distal airspaces of the lungs was measured by the increase in protein concentration of the instilled solution over 1 h. Data are presented as a ratio between final and instilled protein concentration.

Injection:

Article Title: mTORC2 regulates renal tubule sodium uptake by promoting ENaC activity
Article Snippet: After 3 days of acclimatization, vehicle or PP242 solution was injected i.p. at a final dose of 30 mg/kg body weight. .. In the afternoon of day 3 of acclimatization, for studies with amiloride, mice were given 20 mg/l amiloride (Sigma-Aldrich) in their drinking water.

Article Title: Dual activation of CFTR and CLCN2 by lubiprostone in murine nasal epithelia
Article Snippet: Nasal potential difference (NPD) solution “Ringer” saline solution consists of commercial Ringer Injection (1.48 mM NaCl, 2.25 mM CaCl2 ·2H2 O, 4.05 mM KCl, Baxter Healthcare, Deerfield, IL) + 2.4 mM K2 HPO4 , 0.4 mM KH2 PO4 , 1.2 mM MgCl2 ·6H2 O. .. “Amiloride” is Ringer + 100 μM amiloride (Sigma Aldrich).

Article Title: Insulin ameliorates pulmonary edema through the upregulation of epithelial sodium channel via the PI3K/SGK1 pathway in mice with lipopolysaccharide-induced lung injury
Article Snippet: .. Albumin solution containing amiloride (5×10−4 M; Sigma-Aldrich; Merck KGaA) was injected into the alveolar spaces. .. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay Detection of apoptotic cells in the lung epithelium was performed on paraffin-embedded 5-µm lung sections using an in situ cell death detection kit (Roche Diagnostics, Indianapolis, IN, USA) by TUNEL reaction.

Spectrophotometry:

Article Title: Insulin ameliorates pulmonary edema through the upregulation of epithelial sodium channel via the PI3K/SGK1 pathway in mice with lipopolysaccharide-induced lung injury
Article Snippet: Physiological saline solution (5 ml/kg) containing 5% albumin and 0.15 mg/ml Evans blue dye (Sigma-Aldrich; Merck KGaA) was injected into the alveolar spaces at an airway pressure of 7 cm H2 O. Alveolar fluid was aspirated 1 h after instillation, and the concentrations of Evans blue-labeled albumin in the injected and aspirated solutions were measured using a spectrophotometer (BD Bioscience). .. Albumin solution containing amiloride (5×10−4 M; Sigma-Aldrich; Merck KGaA) was injected into the alveolar spaces.

Fluorescence In Situ Hybridization:

Article Title: Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways
Article Snippet: For experiments with the otoprotective agent PROTO1 , larvae were pretreated for 1 hr in embryo medium supplemented with 10 μM PROTO1, then neomycin or gentamicin was added and fish were treated acutely (30 min, followed by 1 hr recovery in fresh EM without PROTO1) or chronically (6 hrs continuous treatment with aminoglycoside and PROTO1). .. For experiments with amiloride and acute aminoglycoside exposure, zebrafish larvae were pretreated for 15 minutes with 1 mM amiloride (Sigma) in embryo medium, transferred to 200 μM neomycin or gentamicin plus 1 mM amiloride for 30 min, washed four times in embryo medium and held in the final rinse for one hour prior to evaluation of hair cells with DASPEI.

Molecular Weight:

Article Title: The influence of mode of delivery, hormonal status and postnatal O2 environment on epithelial sodium channel (ENaC) expression in perinatal guinea-pig lung
Article Snippet: .. To determine the fractional inhibition of lung fluid clearance by amiloride, an excess of amiloride (10−3 M; Sigma) was added to the 5% albumin instillate solution to ensure an appropriate concentration at the alveolar epithelium, since a fraction is protein bound and another fraction rapidly leaves the airspace due to its low molecular weight. .. Clearance of fluid from the distal airspaces of the lungs was measured by the increase in protein concentration of the instilled solution over 1 h. Data are presented as a ratio between final and instilled protein concentration.

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    Millipore amiloride
    Increased transepithelial chloride transport following Dab2 depletion in CFBE41o-WT monolayers CFBE41o-WT cells were transfected with control or Dab2 siRNA oligos. 24 h after transfection, cells were lifted, seeded onto Transwell filters and cultured for additional 4 to 5 days. Dab2 depletion efficiency was > 95%. The short-circuit currents ( I sc ) across the monolayers were measured in Ussing chambers as described in the Experimental Procedures section. A. Representative tracings from control and Dab2 depleted monolayers. Experiments were performed in the setting of a C1 − gradient between the basolateral and apical compartments. Following addition of <t>amiloride</t> (100 μM), 20 μM forskolin (a), 50 μM genistein (b) and 10 μM CFTR Inh -172 were added at times indicated. B. Forskiolin+genistein activated I sc . Δ I sc were calculated as an increase of I sc after forskolin and genistein addition over the base-line currents, n=4.
    Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amiloride/product/Millipore
    Average 95 stars, based on 105 article reviews
    Price from $9.99 to $1999.99
    amiloride - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    75
    Millipore transepithelial amiloride sensitive current
    Measurements of <t>amiloride-sensitive</t> <t>transepithelial</t> current in 2F3 cells. A , transepithelial sodium current decreased with time after treating Xenopus 2F3 cells with 10 μ m cathepsin B inhibitor. B , transepithelial sodium current decreased after
    Transepithelial Amiloride Sensitive Current, supplied by Millipore, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transepithelial amiloride sensitive current/product/Millipore
    Average 75 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transepithelial amiloride sensitive current - by Bioz Stars, 2020-02
    75/100 stars
      Buy from Supplier


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    Increased transepithelial chloride transport following Dab2 depletion in CFBE41o-WT monolayers CFBE41o-WT cells were transfected with control or Dab2 siRNA oligos. 24 h after transfection, cells were lifted, seeded onto Transwell filters and cultured for additional 4 to 5 days. Dab2 depletion efficiency was > 95%. The short-circuit currents ( I sc ) across the monolayers were measured in Ussing chambers as described in the Experimental Procedures section. A. Representative tracings from control and Dab2 depleted monolayers. Experiments were performed in the setting of a C1 − gradient between the basolateral and apical compartments. Following addition of amiloride (100 μM), 20 μM forskolin (a), 50 μM genistein (b) and 10 μM CFTR Inh -172 were added at times indicated. B. Forskiolin+genistein activated I sc . Δ I sc were calculated as an increase of I sc after forskolin and genistein addition over the base-line currents, n=4.

    Journal: The Biochemical journal

    Article Title: Dab2 is a Key Regulator of Endocytosis and Post-endocytic Trafficking of the Cystic Fibrosis Transmembrane Conductance Regulator

    doi: 10.1042/BJ20111566

    Figure Lengend Snippet: Increased transepithelial chloride transport following Dab2 depletion in CFBE41o-WT monolayers CFBE41o-WT cells were transfected with control or Dab2 siRNA oligos. 24 h after transfection, cells were lifted, seeded onto Transwell filters and cultured for additional 4 to 5 days. Dab2 depletion efficiency was > 95%. The short-circuit currents ( I sc ) across the monolayers were measured in Ussing chambers as described in the Experimental Procedures section. A. Representative tracings from control and Dab2 depleted monolayers. Experiments were performed in the setting of a C1 − gradient between the basolateral and apical compartments. Following addition of amiloride (100 μM), 20 μM forskolin (a), 50 μM genistein (b) and 10 μM CFTR Inh -172 were added at times indicated. B. Forskiolin+genistein activated I sc . Δ I sc were calculated as an increase of I sc after forskolin and genistein addition over the base-line currents, n=4.

    Article Snippet: Amiloride (100 μM) was added to block residual Na+ current, followed by the CFTR agonists forskolin (20 μM, Calbiochem) and genistein (50 μM, Sigma Aldridge) as indicated (minimum five-min observation at each concentration).

    Techniques: Transfection, Cell Culture

    Effect of P on AFC response to activators of PKA in normal BALB/c mice. A : effect of 50 μM forskolin (open bars), forskolin + 1 μg/ml 14–22 amide (hatched bars), and forskolin + 1.5 mM amiloride (gray bars) on AFC in BALB/c mice

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Effect of ventilation pressure on alveolar fluid clearance and ?-agonist responses in mice

    doi: 10.1152/ajplung.00096.2009

    Figure Lengend Snippet: Effect of P on AFC response to activators of PKA in normal BALB/c mice. A : effect of 50 μM forskolin (open bars), forskolin + 1 μg/ml 14–22 amide (hatched bars), and forskolin + 1.5 mM amiloride (gray bars) on AFC in BALB/c mice

    Article Snippet: Amiloride, CFTRinh -172 (EMD Biosciences, La Jolla, CA), forskolin, cholera toxin, propranolol, glibenclamide, DPCPX (8-cyclopentyl-1,3-dipropylxanthine; Tocris Bioscience, Ellisville, MO), and ZM241385 (4-(2-[7-Amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol; Tocris) were reconstituted in DMSO.

    Techniques: Mouse Assay

    Addition of amiloride, inhibition of actin polymerization, or PKC inhibition decreased CXCL1 internalization in DIH cells. DIH cells were pre-treated with various inhibitors prior to incubation with 125 I-CXCL1/GRO-α for the specified time points, as described in Materials and Methods . (A) Amiloride significantly inhibited DARC-mediated CXCL1 internalization in DIH cells. * p

    Journal: PLoS ONE

    Article Title: Duffy Antigen Receptor for Chemokines Mediates Chemokine Endocytosis through a Macropinocytosis-Like Process in Endothelial Cells

    doi: 10.1371/journal.pone.0029624

    Figure Lengend Snippet: Addition of amiloride, inhibition of actin polymerization, or PKC inhibition decreased CXCL1 internalization in DIH cells. DIH cells were pre-treated with various inhibitors prior to incubation with 125 I-CXCL1/GRO-α for the specified time points, as described in Materials and Methods . (A) Amiloride significantly inhibited DARC-mediated CXCL1 internalization in DIH cells. * p

    Article Snippet: 2-hydroxypropyl-β-Cyclodextrin (10 mM), dynasore (50 µM), amiloride (3 mM), cytochalasin D (2.5 µM), Gö6976 (1 µM; Calbiochem, Gibbstown, NJ) and PDGF-BB (Millipore, Billerica, MA) (50 ng/ml) were used for endocytosis studies.

    Techniques: Inhibition, Incubation

    Measurements of amiloride-sensitive transepithelial current in 2F3 cells. A , transepithelial sodium current decreased with time after treating Xenopus 2F3 cells with 10 μ m cathepsin B inhibitor. B , transepithelial sodium current decreased after

    Journal: The Journal of Biological Chemistry

    Article Title: Cathepsin B Is Secreted Apically from Xenopus 2F3 Cells and Cleaves the Epithelial Sodium Channel (ENaC) to Increase Its Activity *

    doi: 10.1074/jbc.M111.338574

    Figure Lengend Snippet: Measurements of amiloride-sensitive transepithelial current in 2F3 cells. A , transepithelial sodium current decreased with time after treating Xenopus 2F3 cells with 10 μ m cathepsin B inhibitor. B , transepithelial sodium current decreased after

    Article Snippet: The voltage and resistance across confluent monolayers were measured with an epithelial Voltohmeter (EVOM; World Precision Instrument, Sarasota, FL), and transepithelial amiloride-sensitive current was calculated per unit area according to Ohm's law before and after application of 10 μ m cathepsin B inhibitor (CA-74) (Calbiochem) or vehicle dimethyl sulfoxide (DMSO) (Sigma) at time points between 5 and 60 min.

    Techniques: