amiloride  (Millipore)

 
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  • 97
    Name:
    Amiloride hydrochloride hydrate
    Description:

    Catalog Number:
    A7410
    Price:
    None
    Applications:
    Amiloride is a selective T-type calcium channel blocker, an epithelial sodium channel blocker and a selective inhibitor of urokinase plasminogen activator (uPA). Amiloride has been used in a study to develop an alternative treatment for constipation.
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    Structured Review

    Millipore amiloride
    Amiloride hydrochloride hydrate

    https://www.bioz.com/result/amiloride/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    amiloride - by Bioz Stars, 2021-06
    97/100 stars

    Images

    1) Product Images from "CHBPR-Angiotensin II stimulates renin in inner medullary collecting duct cells via PKC and independent of ENaC and mineralocorticoid receptor activity"

    Article Title: CHBPR-Angiotensin II stimulates renin in inner medullary collecting duct cells via PKC and independent of ENaC and mineralocorticoid receptor activity

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.110.165902

    A. Renin mRNA levels in medullary tissues in each group. B. Urinary renin content was increased in Ang II infused rats and Ang II infused rats treated with amiloride. * P
    Figure Legend Snippet: A. Renin mRNA levels in medullary tissues in each group. B. Urinary renin content was increased in Ang II infused rats and Ang II infused rats treated with amiloride. * P

    Techniques Used:

    A. Progression of systolic blood pressure in control rats (Sham operated), Ang II-infused and Ang II-infused treated with amiloride. Regression line curve (R 2 ) is shown for each group (n=10). B. Ang II content in kidney cortex and medulla. * P
    Figure Legend Snippet: A. Progression of systolic blood pressure in control rats (Sham operated), Ang II-infused and Ang II-infused treated with amiloride. Regression line curve (R 2 ) is shown for each group (n=10). B. Ang II content in kidney cortex and medulla. * P

    Techniques Used:

    2) Product Images from "Cav3-type α1T calcium channels mediate transient calcium currents that regulate repetitive firing in Drosophila antennal lobe PNs"

    Article Title: Cav3-type α1T calcium channels mediate transient calcium currents that regulate repetitive firing in Drosophila antennal lobe PNs

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00368.2013

    Isolation of amiloride-sensitive transient calcium currents by prepulse inactivation. A : transient (T) current evoked by a test step to −5 mV from a prepulse of −80 mV (PP −80) is blocked by prepulse to −10 mV (PP −10).
    Figure Legend Snippet: Isolation of amiloride-sensitive transient calcium currents by prepulse inactivation. A : transient (T) current evoked by a test step to −5 mV from a prepulse of −80 mV (PP −80) is blocked by prepulse to −10 mV (PP −10).

    Techniques Used: Isolation

    Transient calcium currents in projections neurons (PNs) are amiloride sensitive. A : isolated calcium currents in a PN in brain of wild-type adult with both transient and sustained components. Currents elicited by a series of depolarizing voltage steps
    Figure Legend Snippet: Transient calcium currents in projections neurons (PNs) are amiloride sensitive. A : isolated calcium currents in a PN in brain of wild-type adult with both transient and sustained components. Currents elicited by a series of depolarizing voltage steps

    Techniques Used: Isolation

    3) Product Images from "Acid-sensing ion channels in acidosis-induced injury of human brain neurons"

    Article Title: Acid-sensing ion channels in acidosis-induced injury of human brain neurons

    Journal: Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism

    doi: 10.1038/jcbfm.2010.30

    Pharmacology of acid-sensing ion channels (ASICs) in human cortical neurons. ( A and B ) Representative current traces and summary data showing the blockade of ASIC currents by amiloride. ( C and D ) Representative current traces and summary data showing
    Figure Legend Snippet: Pharmacology of acid-sensing ion channels (ASICs) in human cortical neurons. ( A and B ) Representative current traces and summary data showing the blockade of ASIC currents by amiloride. ( C and D ) Representative current traces and summary data showing

    Techniques Used:

    4) Product Images from "Dural afferents express acid-sensing ion channels: a role for decreased meningeal pH in migraine headache"

    Article Title: Dural afferents express acid-sensing ion channels: a role for decreased meningeal pH in migraine headache

    Journal: Pain

    doi: 10.1016/j.pain.2010.09.036

    Amiloride blockade of pH 6.0 evoked currents in dural afferents. A–C) pH was stepped from 7.4 to 6.0 for 1s or 5s every 20s. pH 6.0 evoked current in a representative dural afferent is reversibly blocked by 1 mM amiloride (A) but not 10 μM
    Figure Legend Snippet: Amiloride blockade of pH 6.0 evoked currents in dural afferents. A–C) pH was stepped from 7.4 to 6.0 for 1s or 5s every 20s. pH 6.0 evoked current in a representative dural afferent is reversibly blocked by 1 mM amiloride (A) but not 10 μM

    Techniques Used:

    Amiloride exhibits paradoxical effect on higher pH. A) pH 6.9 evoked current in a representative dural afferent is blocked by 1 mM amiloride. B) The percentage of pH 6.9 evoked peak current amplitude blocked by 1 mM amiloride (n = 9), 10 μM capsazepine
    Figure Legend Snippet: Amiloride exhibits paradoxical effect on higher pH. A) pH 6.9 evoked current in a representative dural afferent is blocked by 1 mM amiloride. B) The percentage of pH 6.9 evoked peak current amplitude blocked by 1 mM amiloride (n = 9), 10 μM capsazepine

    Techniques Used:

    5) Product Images from "Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes"

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    Journal: British Journal of Pharmacology

    doi: 10.1038/sj.bjp.0703520

    Effect of genistein on ion transport in rectal tissues from non-CF and CF individuals. (A) Concentration response curve for the effect of genistein (in the presence of 10 μmol l −1 indomethacin; basolateral side) on non-CF rectal mucosa (EC 50 =35 μmol l −1 ). (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF rectal mucosa. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either(i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). # Indicates statistical significance for the effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect of genistein in the absence of cyclic AMP activation, in the presence of forskolin and in the presence of IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).
    Figure Legend Snippet: Effect of genistein on ion transport in rectal tissues from non-CF and CF individuals. (A) Concentration response curve for the effect of genistein (in the presence of 10 μmol l −1 indomethacin; basolateral side) on non-CF rectal mucosa (EC 50 =35 μmol l −1 ). (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF rectal mucosa. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either(i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). # Indicates statistical significance for the effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect of genistein in the absence of cyclic AMP activation, in the presence of forskolin and in the presence of IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Techniques Used: Concentration Assay, Activation Assay

    Effects of genistein on cholinergic secretion after deactivating (indomethacin) or activating (forskolin/IBMX) CFTR. Original recordings of V te obtained from rectal biopsies from (A) non-CF and (B) CF subjects. All experiments were performed in the presence of indomethacin (10 μmol l −1 ; basolateral side) and amiloride (10 μmol l −1 ; mucosal side). Cholinergic stimulation was induced by carbachol (CCH, 100 μmol l −1 ; basolateral side). (C) Summary of the CCH induced I sc in CF and non-CF rectal biopsies in the absence and presence of genistein (50 μmol l −1 mucosal side) and dependence on stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). #Indicates significant difference for CCH induced I sc in the presence of IBMX/forskolin compared to indomethacin (paired t -test). * Indicates statistical significance for the effect of genistein on CCH induced I sc in the presence of indomethacin and IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of genistein on CCH induced I sc in experiments obtained from non-CF and CF tissues (unpaired t -test). (Number of subjects).
    Figure Legend Snippet: Effects of genistein on cholinergic secretion after deactivating (indomethacin) or activating (forskolin/IBMX) CFTR. Original recordings of V te obtained from rectal biopsies from (A) non-CF and (B) CF subjects. All experiments were performed in the presence of indomethacin (10 μmol l −1 ; basolateral side) and amiloride (10 μmol l −1 ; mucosal side). Cholinergic stimulation was induced by carbachol (CCH, 100 μmol l −1 ; basolateral side). (C) Summary of the CCH induced I sc in CF and non-CF rectal biopsies in the absence and presence of genistein (50 μmol l −1 mucosal side) and dependence on stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). #Indicates significant difference for CCH induced I sc in the presence of IBMX/forskolin compared to indomethacin (paired t -test). * Indicates statistical significance for the effect of genistein on CCH induced I sc in the presence of indomethacin and IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of genistein on CCH induced I sc in experiments obtained from non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Techniques Used:

    Effect of genistein on ion transport in nasal epithelia from non-CF and CF subjects. (A) Concentration response curve for the effect of genistein (under resting conditions) in non-CF respiratory epithelia, showing an EC 50 of about 3.8 μmol l −1 . An inhibitory effect was observed at concentrations higher than 50 μmol l −1 . (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF nasal epithelia. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either (i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides). #Significant effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect genistein compared to control and in the presence of forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).
    Figure Legend Snippet: Effect of genistein on ion transport in nasal epithelia from non-CF and CF subjects. (A) Concentration response curve for the effect of genistein (under resting conditions) in non-CF respiratory epithelia, showing an EC 50 of about 3.8 μmol l −1 . An inhibitory effect was observed at concentrations higher than 50 μmol l −1 . (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF nasal epithelia. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either (i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides). #Significant effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect genistein compared to control and in the presence of forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Techniques Used: Concentration Assay

    (A) Summary of short circuit currents (I sc ) in rectal epithelia from non-CF and CF individuals under resting conditions, after incubation with indomethacin (10 μmol l −1 ; basolateral side), after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). * Indicates statistical significance for the effects of indomethacin, amiloride and IBMX/forskolin in non-CF or CF (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of indomethacin, amiloride and IBMX/forskolin obtained in experiments from rectal tissues of non-CF and CF subjects (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human rectal mucosa. The effects of genistein were examined in both absence and presence of IBMX and forskolin and experiments were carried out in the presence of indomethacin and amiloride. R te was continuously determined from the V te downward deflections obtained by pulsed current injection.
    Figure Legend Snippet: (A) Summary of short circuit currents (I sc ) in rectal epithelia from non-CF and CF individuals under resting conditions, after incubation with indomethacin (10 μmol l −1 ; basolateral side), after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). * Indicates statistical significance for the effects of indomethacin, amiloride and IBMX/forskolin in non-CF or CF (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of indomethacin, amiloride and IBMX/forskolin obtained in experiments from rectal tissues of non-CF and CF subjects (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human rectal mucosa. The effects of genistein were examined in both absence and presence of IBMX and forskolin and experiments were carried out in the presence of indomethacin and amiloride. R te was continuously determined from the V te downward deflections obtained by pulsed current injection.

    Techniques Used: Incubation, Blocking Assay, Activation Assay, Injection

    (A) Summary of short circuit currents (I sc ) under resting conditions, after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides) in human nasal epithelia from non-CF and CF individuals. * Indicates statistical significance for the effects of amiloride and IBMX/forskolin (paired t -test). §Indicate statistical significance for the basal I sc and the effects of amiloride and IBMX/forskolin when compared to experiments obtained from non-CF tissues (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human nasal tissues. Epithelial Na + conductance was blocked by amiloride and effects of genistein were examined in both absence and presence of IBMX and forskolin. R te was determined continuously from the V te downward deflections obtained by pulsed current injection.
    Figure Legend Snippet: (A) Summary of short circuit currents (I sc ) under resting conditions, after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides) in human nasal epithelia from non-CF and CF individuals. * Indicates statistical significance for the effects of amiloride and IBMX/forskolin (paired t -test). §Indicate statistical significance for the basal I sc and the effects of amiloride and IBMX/forskolin when compared to experiments obtained from non-CF tissues (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human nasal tissues. Epithelial Na + conductance was blocked by amiloride and effects of genistein were examined in both absence and presence of IBMX and forskolin. R te was determined continuously from the V te downward deflections obtained by pulsed current injection.

    Techniques Used: Blocking Assay, Activation Assay, Injection

    6) Product Images from "Effects of amiloride, benzamil, and alterations in extracellular Na+ on the rat afferent arteriole and its myogenic response"

    Article Title: Effects of amiloride, benzamil, and alterations in extracellular Na+ on the rat afferent arteriole and its myogenic response

    Journal:

    doi: 10.1152/ajprenal.00200.2007

    Original tracings ( left ) and mean data showing effects of 10 μmol/l benzamil ( n = 4), 3 μmol/l amiloride ( A and B , n = 3), and reducing [Na + ] o from 140 to 100 mmol/l ( C and D , n = 4) on the
    Figure Legend Snippet: Original tracings ( left ) and mean data showing effects of 10 μmol/l benzamil ( n = 4), 3 μmol/l amiloride ( A and B , n = 3), and reducing [Na + ] o from 140 to 100 mmol/l ( C and D , n = 4) on the

    Techniques Used:

    Amiloride had no significant effect on the myogenic response of the afferent arteriole to changes in renal arterial pressure ( n = 5).
    Figure Legend Snippet: Amiloride had no significant effect on the myogenic response of the afferent arteriole to changes in renal arterial pressure ( n = 5).

    Techniques Used:

    7) Product Images from "Dynamic Effects of Hg2+-induced Changes in Cell Volume"

    Article Title: Dynamic Effects of Hg2+-induced Changes in Cell Volume

    Journal:

    doi: 10.1007/s12013-008-9010-y

    Effect of amiloride on Hg 2+ effects. ( a ) 10 μM amiloride and ( b ) 100 μM amiloride. Amiloride in high enough concentrations to block ENaC and NHE partially blocked the second swelling
    Figure Legend Snippet: Effect of amiloride on Hg 2+ effects. ( a ) 10 μM amiloride and ( b ) 100 μM amiloride. Amiloride in high enough concentrations to block ENaC and NHE partially blocked the second swelling

    Techniques Used: Blocking Assay

    Intracellular pH following 100 μM Hg 2+ treatment. ( a ) In isotonic media, Hg 2+ induces a sudden drop in pH that recovers within 2 min. ( b ) Hypotonic media introduces a similar response to isotonic media, but amiloride at 1 mM that eliminates the
    Figure Legend Snippet: Intracellular pH following 100 μM Hg 2+ treatment. ( a ) In isotonic media, Hg 2+ induces a sudden drop in pH that recovers within 2 min. ( b ) Hypotonic media introduces a similar response to isotonic media, but amiloride at 1 mM that eliminates the

    Techniques Used:

    8) Product Images from "Aldosterone acts via an ATP autocrine/paracrine system: The Edelman ATP hypothesis revisited"

    Article Title: Aldosterone acts via an ATP autocrine/paracrine system: The Edelman ATP hypothesis revisited

    Journal:

    doi: 10.1073/pnas.0507008102

    Effect of osmotic stress and aldosterone on G t and the topography of A6 cell monolayer. ( A ) Typical time-dependent effect of basolateral hypoosmotic stress on the amiloride-sensitive G t ENaC of A6 monolayer. ( B ) SICM images of a selected area of A6 monolayer
    Figure Legend Snippet: Effect of osmotic stress and aldosterone on G t and the topography of A6 cell monolayer. ( A ) Typical time-dependent effect of basolateral hypoosmotic stress on the amiloride-sensitive G t ENaC of A6 monolayer. ( B ) SICM images of a selected area of A6 monolayer

    Techniques Used:

    Aldosterone induces ATP release and changes the morphology of A6 cells. ( A ) Effect of control ( n = 8), aldosterone ( n = 8), and basolateral hypotonic stress ( n = 7) on the amiloride-sensitive transepithelial electrical conductance (G t ENaC ) of A6 cell
    Figure Legend Snippet: Aldosterone induces ATP release and changes the morphology of A6 cells. ( A ) Effect of control ( n = 8), aldosterone ( n = 8), and basolateral hypotonic stress ( n = 7) on the amiloride-sensitive transepithelial electrical conductance (G t ENaC ) of A6 cell

    Techniques Used:

    9) Product Images from "Regulation of the Epithelial Na+ Channel by the Protein Kinase CK2 * Channel by the Protein Kinase CK2 * S⃞"

    Article Title: Regulation of the Epithelial Na+ Channel by the Protein Kinase CK2 * Channel by the Protein Kinase CK2 * S⃞

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M704532200

    CK2 activates ENaC in Xenopus oocytes. A , current recording from a Xenopus oocyte expressing αβγ-ENaC and effects of amiloride (10 μ m ) and TBB (10 μ m ). Oocytes were voltage-clamped from -90 mV to +10 mV in steps of 10 mV, and the resulting currents were recorded. B , summary of the effects of amiloride and TBB on whole cell conductance in ENaC-expressing oocytes. C , summary of the change of amiloride-sensitive conductance ( G amil ) after injection of water, the CK2 inhibitor poly(E:Y) (50 μ m ) and CK2 activator poly(K) (50 μ m ), respectively. D , current recording of an ENaC-expressing oocyte and the effects of amiloride ( A , 10 μ m ) and okadaic acid (100 n m ). E , summary of the effect of okadaic acid on the amiloride-sensitive whole cell conductance measured in oocytes. The asterisk ( * ) indicates significant effects (paired t -test). The number sign (#) indicates a significant difference for the effects of amiloride before and after incubation with TBB (paired t -test, 6-25 experiments for each series).
    Figure Legend Snippet: CK2 activates ENaC in Xenopus oocytes. A , current recording from a Xenopus oocyte expressing αβγ-ENaC and effects of amiloride (10 μ m ) and TBB (10 μ m ). Oocytes were voltage-clamped from -90 mV to +10 mV in steps of 10 mV, and the resulting currents were recorded. B , summary of the effects of amiloride and TBB on whole cell conductance in ENaC-expressing oocytes. C , summary of the change of amiloride-sensitive conductance ( G amil ) after injection of water, the CK2 inhibitor poly(E:Y) (50 μ m ) and CK2 activator poly(K) (50 μ m ), respectively. D , current recording of an ENaC-expressing oocyte and the effects of amiloride ( A , 10 μ m ) and okadaic acid (100 n m ). E , summary of the effect of okadaic acid on the amiloride-sensitive whole cell conductance measured in oocytes. The asterisk ( * ) indicates significant effects (paired t -test). The number sign (#) indicates a significant difference for the effects of amiloride before and after incubation with TBB (paired t -test, 6-25 experiments for each series).

    Techniques Used: Expressing, Injection, Incubation

    Elimination of CK2 phosphorylation sites on ENaC inhibits channel activity. A , current recording from a Xenopus oocyte expressing αβγ-ENaC and effects of amiloride (10μ m ) and TBB (10μ m ). Oocytes were voltage-clamped from -90 mV to +10 mV in steps of 10 mV, and the resulting currents were recorded. B , current recording from a Xenopus oocyte expressing αβ S631A γ T559A -ENaC and effects of amiloride (10 μ m ) and TBB (10 μ m ). C and D , summaries of the effects of amiloride and TBB on G amil generated by αβγ-ENaC and αβ S631A γ T559A -ENaC. E , comparison of G amil produced by wt-(αβγ)-, single mutants (αβ S631A γ, αβγ T559A )-, and a doublemutant (αβ S631A γ T559A )-ENaC. F , summaries of G amil produced by dimeric wt-(αβ, αγ)- and mutant (αβ S631A , αγ T559 )-ENaC channels. The asterisk ( * ) and number sign (#) indicate a significant difference (paired t -test, 13-25 experiments for each series).
    Figure Legend Snippet: Elimination of CK2 phosphorylation sites on ENaC inhibits channel activity. A , current recording from a Xenopus oocyte expressing αβγ-ENaC and effects of amiloride (10μ m ) and TBB (10μ m ). Oocytes were voltage-clamped from -90 mV to +10 mV in steps of 10 mV, and the resulting currents were recorded. B , current recording from a Xenopus oocyte expressing αβ S631A γ T559A -ENaC and effects of amiloride (10 μ m ) and TBB (10 μ m ). C and D , summaries of the effects of amiloride and TBB on G amil generated by αβγ-ENaC and αβ S631A γ T559A -ENaC. E , comparison of G amil produced by wt-(αβγ)-, single mutants (αβ S631A γ, αβγ T559A )-, and a doublemutant (αβ S631A γ T559A )-ENaC. F , summaries of G amil produced by dimeric wt-(αβ, αγ)- and mutant (αβ S631A , αγ T559 )-ENaC channels. The asterisk ( * ) and number sign (#) indicate a significant difference (paired t -test, 13-25 experiments for each series).

    Techniques Used: Activity Assay, Expressing, Generated, Produced, Mutagenesis

    CK2 is essential for ENaC activity and antagonizes the inhibitory effect of Nedd4-2 on ENaC. A , summary of α-ENaC membrane expression and G amil after 40 h. TBB inhibited membrane expression of α-ENaC via single mutants (αβ S631A γ,αβγ T559A ) but not that of the double mutant (αβ S631A γ T559A ). G amil was largely reduced for all mutants, and G amil produced by the double mutant was no longer inhibited by TBB. Dashed lines indicate membrane expression and G amil of wt-ENaC. B , whole cell conductances relative to wt-ENaC. A mutation in the PY motif (Y618A) of β-ENaC increased Na + conductance, and S631A no longer inhibited ENaC conductance. The Grk2 mutant S633A inhibited ENaC, but not as a double mutant S631A/S633A. C , inhibition of xNedd4-2 expression by siRNA-xNedd4-2 but not scrambled siRNA. The abundant poly(A)-binding protein indicates equal loading. Summary of ENaC whole cell conductances measured in the absence or presence of siRNA-xNedd4-2 or scrambled siRNA (see “Materials and Methods”). D , summary of the amiloride-sensitive short-circuit current and effects of TBB (10 μ m ) in control M1 cells and M1 cell treated with scrambled RNAi, mNedd4-2-RNAi, and mCK2-RNAi (see “Materials and Methods”). The asterisk ( * ) indicates significant effects of TBB (paired t -tests). The number sign (#) indicates a significant difference compared with control (unpaired t -test, 6-24 experiments for each series).
    Figure Legend Snippet: CK2 is essential for ENaC activity and antagonizes the inhibitory effect of Nedd4-2 on ENaC. A , summary of α-ENaC membrane expression and G amil after 40 h. TBB inhibited membrane expression of α-ENaC via single mutants (αβ S631A γ,αβγ T559A ) but not that of the double mutant (αβ S631A γ T559A ). G amil was largely reduced for all mutants, and G amil produced by the double mutant was no longer inhibited by TBB. Dashed lines indicate membrane expression and G amil of wt-ENaC. B , whole cell conductances relative to wt-ENaC. A mutation in the PY motif (Y618A) of β-ENaC increased Na + conductance, and S631A no longer inhibited ENaC conductance. The Grk2 mutant S633A inhibited ENaC, but not as a double mutant S631A/S633A. C , inhibition of xNedd4-2 expression by siRNA-xNedd4-2 but not scrambled siRNA. The abundant poly(A)-binding protein indicates equal loading. Summary of ENaC whole cell conductances measured in the absence or presence of siRNA-xNedd4-2 or scrambled siRNA (see “Materials and Methods”). D , summary of the amiloride-sensitive short-circuit current and effects of TBB (10 μ m ) in control M1 cells and M1 cell treated with scrambled RNAi, mNedd4-2-RNAi, and mCK2-RNAi (see “Materials and Methods”). The asterisk ( * ) indicates significant effects of TBB (paired t -tests). The number sign (#) indicates a significant difference compared with control (unpaired t -test, 6-24 experiments for each series).

    Techniques Used: Activity Assay, Expressing, Mutagenesis, Produced, Inhibition, Binding Assay

    CK2 activates ENaC in native epithelia and in epithelial cells. Original Ussing chamber recordings of the transepithelial voltages V te detected in mouse trachea ( A ), mouse colon ( C ), and M1 cells ( E ). Effects of amiloride ( A , 10 μ m ) and the CK2 inhibitor TBB (10 μ m ). Concentration-dependence of the effects of TBB on amiloride-sensitive transport in trachea ( B ), colon ( D ), and M1 cells ( F ). The asterisk ( * ) indicates significant effects of TBB (paired t -tests, number of experiments: 9-13 for each series).
    Figure Legend Snippet: CK2 activates ENaC in native epithelia and in epithelial cells. Original Ussing chamber recordings of the transepithelial voltages V te detected in mouse trachea ( A ), mouse colon ( C ), and M1 cells ( E ). Effects of amiloride ( A , 10 μ m ) and the CK2 inhibitor TBB (10 μ m ). Concentration-dependence of the effects of TBB on amiloride-sensitive transport in trachea ( B ), colon ( D ), and M1 cells ( F ). The asterisk ( * ) indicates significant effects of TBB (paired t -tests, number of experiments: 9-13 for each series).

    Techniques Used: Concentration Assay

    10) Product Images from "Physiological levels of lipoxin A4 inhibit ENaC and restore airway surface liquid height in cystic fibrosis bronchial epithelium"

    Article Title: Physiological levels of lipoxin A4 inhibit ENaC and restore airway surface liquid height in cystic fibrosis bronchial epithelium

    Journal: Physiological Reports

    doi: 10.14814/phy2.12093

    Effect of LXA 4 on amiloride‐sensitive ion transport and ASL height regulation. Response of the amiloride‐sensitive current to LXA 4 (1 nmol/L) pretreatment in NuLi‐1 (A) CuFi‐1 (B) monolayers. Effect of apical amiloride (1 μ mol/L, 15 min treatment) application on the LXA 4 mediated increase in ASL height in NuLi‐1 (C) CuFi‐1 (D) monolayers (* P
    Figure Legend Snippet: Effect of LXA 4 on amiloride‐sensitive ion transport and ASL height regulation. Response of the amiloride‐sensitive current to LXA 4 (1 nmol/L) pretreatment in NuLi‐1 (A) CuFi‐1 (B) monolayers. Effect of apical amiloride (1 μ mol/L, 15 min treatment) application on the LXA 4 mediated increase in ASL height in NuLi‐1 (C) CuFi‐1 (D) monolayers (* P

    Techniques Used:

    11) Product Images from "Effect of inhibitors of endocytosis and NF-kB signal pathway on folate-conjugated nanoparticle endocytosis by rat Kupffer cells"

    Article Title: Effect of inhibitors of endocytosis and NF-kB signal pathway on folate-conjugated nanoparticle endocytosis by rat Kupffer cells

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S141407

    Dose-dependent inhibition of KC cytokines by compound inhibitors. Notes: ( A ) The dose-dependent inhibition effect of compound inhibitors on cytokine TNF-α. ( B ) The dose-dependent inhibition effect of compound inhibitors on cytokine IL-1β. ( C ) The dose-dependent inhibition effect of compound inhibitors on cytokine IL-6. KCs were incubated with different endocytosis compound inhibitors for 24 h prior to addition of 200 µg/mL FPA/EPI NPs, and the cytokine inhibition was quantified by fluorometry compared with untreated controls. Data are represented as mean ± SD of five determinations. Abbreviations: AMR, amiloride; Col, colchicine; CPZ, chlorpromazine; FPA/EPI NPs, epirubicin-loaded folic acid-conjugated pullulan acetate nanoparticles; IL, interleukin; KCs, Kupffer cells; NY, nystatin; PDTC, pyrrolidine dithiocarbamate; TNF-α, tumor necrosis factor.
    Figure Legend Snippet: Dose-dependent inhibition of KC cytokines by compound inhibitors. Notes: ( A ) The dose-dependent inhibition effect of compound inhibitors on cytokine TNF-α. ( B ) The dose-dependent inhibition effect of compound inhibitors on cytokine IL-1β. ( C ) The dose-dependent inhibition effect of compound inhibitors on cytokine IL-6. KCs were incubated with different endocytosis compound inhibitors for 24 h prior to addition of 200 µg/mL FPA/EPI NPs, and the cytokine inhibition was quantified by fluorometry compared with untreated controls. Data are represented as mean ± SD of five determinations. Abbreviations: AMR, amiloride; Col, colchicine; CPZ, chlorpromazine; FPA/EPI NPs, epirubicin-loaded folic acid-conjugated pullulan acetate nanoparticles; IL, interleukin; KCs, Kupffer cells; NY, nystatin; PDTC, pyrrolidine dithiocarbamate; TNF-α, tumor necrosis factor.

    Techniques Used: Inhibition, Incubation

    12) Product Images from "Modulation the alternative splicing of GLA (IVS4+919G > A) in Fabry disease"

    Article Title: Modulation the alternative splicing of GLA (IVS4+919G > A) in Fabry disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0175929

    Effects of amiloride on the regulation of GLA (IVS4 + 919G > A) splicing. Cells were treated with different concentrations of amiloride for 24 hours and then harvested for RT-PCR analysis (A), or Western blot analysis (B,D). Actin and Histone H3 were used as internal standards. (C) The result of enzyme activity assay from FD cells after the treatment with or without amiloride for 24 hours. Data were presented as the mean ± standard deviation from three independent experiments. Asterisk represents significant difference ( p -value
    Figure Legend Snippet: Effects of amiloride on the regulation of GLA (IVS4 + 919G > A) splicing. Cells were treated with different concentrations of amiloride for 24 hours and then harvested for RT-PCR analysis (A), or Western blot analysis (B,D). Actin and Histone H3 were used as internal standards. (C) The result of enzyme activity assay from FD cells after the treatment with or without amiloride for 24 hours. Data were presented as the mean ± standard deviation from three independent experiments. Asterisk represents significant difference ( p -value

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Western Blot, Enzyme Activity Assay, Standard Deviation

    Alterations in histone modification patterns after amiloride treatment. ChIP assays were performed with antibodies to the indicated histone modifications on the cryptic exon area in Int4 of GLA in normal cells or in FD cells treated with or without amiloride. Results were expressed as a fraction of histone H3 after normalization to input values and presented as a mean values ± standard deviation from three independent experiments. Asterisk represents significant difference ( p -value
    Figure Legend Snippet: Alterations in histone modification patterns after amiloride treatment. ChIP assays were performed with antibodies to the indicated histone modifications on the cryptic exon area in Int4 of GLA in normal cells or in FD cells treated with or without amiloride. Results were expressed as a fraction of histone H3 after normalization to input values and presented as a mean values ± standard deviation from three independent experiments. Asterisk represents significant difference ( p -value

    Techniques Used: Modification, Chromatin Immunoprecipitation, Standard Deviation

    13) Product Images from "Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides"

    Article Title: Enhancing gene delivery of adeno-associated viruses by cell-permeable peptides

    Journal: Molecular Therapy. Methods & Clinical Development

    doi: 10.1038/mtm.2013.12

    Entry mechanisms of adeno-associated virus type 2 (AAV2)-cell-permeable peptides (CPP) complexes. (a) The internalization of AAV2 or AAV2-CPP complexes in cells after 30 minutes incubation at 37 or 4 °C. (b) The effect of inhibitory drugs on viral transduction of cells infected by AAV2 or AAV2-CPP complexes. HEK293T cells were preincubated with chlorpromazine (CPZ, 30 nmol/l), filipin (15 nmol/l), or Amiloride (1 mmol/l) for 30 minutes at 37 °C. Treated and untreated cells were spin-infected with the AAV2 alone or complexes of AAV2 with peptides (final concentration: 200 μmol/l) for 90 minutes in the presence of drugs. Green flourescent protein (GFP) expression was analyzed 2 days after infection. All error bars represent the standard deviation of the mean from triplicate experiments. Asterisks indicate statistical significance compared to the no drug treatment group (* P
    Figure Legend Snippet: Entry mechanisms of adeno-associated virus type 2 (AAV2)-cell-permeable peptides (CPP) complexes. (a) The internalization of AAV2 or AAV2-CPP complexes in cells after 30 minutes incubation at 37 or 4 °C. (b) The effect of inhibitory drugs on viral transduction of cells infected by AAV2 or AAV2-CPP complexes. HEK293T cells were preincubated with chlorpromazine (CPZ, 30 nmol/l), filipin (15 nmol/l), or Amiloride (1 mmol/l) for 30 minutes at 37 °C. Treated and untreated cells were spin-infected with the AAV2 alone or complexes of AAV2 with peptides (final concentration: 200 μmol/l) for 90 minutes in the presence of drugs. Green flourescent protein (GFP) expression was analyzed 2 days after infection. All error bars represent the standard deviation of the mean from triplicate experiments. Asterisks indicate statistical significance compared to the no drug treatment group (* P

    Techniques Used: Conditioned Place Preference, Incubation, Transduction, Infection, Concentration Assay, Expressing, Standard Deviation

    14) Product Images from "Basolateral Na+-H+ exchanger-1 in rat taste receptor cells is involved in neural adaptation to acidic stimuli"

    Article Title: Basolateral Na+-H+ exchanger-1 in rat taste receptor cells is involved in neural adaptation to acidic stimuli

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2003.057745

    Proposed model for the mechanism of neural adaptation to acidic stimuli Our data suggest that the basolateral NHE-1 is involved in neural adaptation during sour taste transduction. Acidic stimuli applied to the lingual surface induce a decrease in TRC pH i as a result of the entry of acid equivalents across the apical cell membranes of TRCs. The apical membranes of TRCs exhibit an amiloride- and Ca 2+ -insensitive, cAMP-dependent H + conductive pathway for strong acids (purple). In contrast, weak organic acids permeate across the apical membranes passively as undissociated molecules. The pH i recovery occurs, in part, due to the presence of zoniporide- and cariporide-sensitive basolateral NHE-1. Although NHE-3 is present in the apical membranes of TRCs, under our experimental conditions the NHE-3 seems to be quiescent, and does not participate in pH i regulation in TRCs (green). In the model the pathways for the apical entry and the basolateral exit of acid equivalents are shown in a single TRC. However, within a taste bud, TRCs are heterogeneous, and it is quite likely that not all of the above components are present in all cells. The favourable Na + gradient for the Na + –H + exchangers is maintained by the basolateral Na + –K + -ATPase. An increase in TRC [Ca 2+ ] i induced by ionomycin, activates basolateral NHE-1, increasing the neural adaptation in CT responses to acidic stimuli. Zoniporide, a specific blocker of NHE-1, increased the magnitude of the CT responses to acidic stimulation under control conditions, and completely reversed the ionomycin-induced increase in neural adaptation to acidic stimuli. We conclude that during sour taste transduction the basolateral NHE-1 plays an important role in determining the adaptation (tonic) phase of the CT taste nerve responses to acidic stimuli.
    Figure Legend Snippet: Proposed model for the mechanism of neural adaptation to acidic stimuli Our data suggest that the basolateral NHE-1 is involved in neural adaptation during sour taste transduction. Acidic stimuli applied to the lingual surface induce a decrease in TRC pH i as a result of the entry of acid equivalents across the apical cell membranes of TRCs. The apical membranes of TRCs exhibit an amiloride- and Ca 2+ -insensitive, cAMP-dependent H + conductive pathway for strong acids (purple). In contrast, weak organic acids permeate across the apical membranes passively as undissociated molecules. The pH i recovery occurs, in part, due to the presence of zoniporide- and cariporide-sensitive basolateral NHE-1. Although NHE-3 is present in the apical membranes of TRCs, under our experimental conditions the NHE-3 seems to be quiescent, and does not participate in pH i regulation in TRCs (green). In the model the pathways for the apical entry and the basolateral exit of acid equivalents are shown in a single TRC. However, within a taste bud, TRCs are heterogeneous, and it is quite likely that not all of the above components are present in all cells. The favourable Na + gradient for the Na + –H + exchangers is maintained by the basolateral Na + –K + -ATPase. An increase in TRC [Ca 2+ ] i induced by ionomycin, activates basolateral NHE-1, increasing the neural adaptation in CT responses to acidic stimuli. Zoniporide, a specific blocker of NHE-1, increased the magnitude of the CT responses to acidic stimulation under control conditions, and completely reversed the ionomycin-induced increase in neural adaptation to acidic stimuli. We conclude that during sour taste transduction the basolateral NHE-1 plays an important role in determining the adaptation (tonic) phase of the CT taste nerve responses to acidic stimuli.

    Techniques Used: Transduction

    15) Product Images from "Synaptic Acidification Enhances GABAA Signaling"

    Article Title: Synaptic Acidification Enhances GABAA Signaling

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.6364-09.2010

    NHE inhibition reduces mIPSC amplitude and charge transfer. Series A, 20 µM amiloride (AML) was used to block the NHE. A1, Raw trace sequence from one CGC illustrates the effects of amiloride, protons, amiloride + protons, and increased buffering on mIPSCs. A2 left , averaged traces from one CGC, note that inhibitory effects of amiloride are negated at pH 6.8. A2 right , averaged traces from another CGC illustrate the non-additive effects of amiloride and increased buffering on mIPSCs, scale bar indicates 40 pA and 10 ms. A3 , Effect of experimental amiloride conditions on charge transfer. Series B, NHE block was achieved by substituting extracellular NaCl with LiCl. B1, Raw trace sequence from one CGC illustrates the effects of LiCl replacement, increased buffering capacity and a combination of the two conditions. B2 , averaged traces from one CGC under the conditions specified. B3 , effects of lithium on charge transfer. n=9 to 19 neurons in amiloride experiments, n=3 in lithium experiments. ★ p
    Figure Legend Snippet: NHE inhibition reduces mIPSC amplitude and charge transfer. Series A, 20 µM amiloride (AML) was used to block the NHE. A1, Raw trace sequence from one CGC illustrates the effects of amiloride, protons, amiloride + protons, and increased buffering on mIPSCs. A2 left , averaged traces from one CGC, note that inhibitory effects of amiloride are negated at pH 6.8. A2 right , averaged traces from another CGC illustrate the non-additive effects of amiloride and increased buffering on mIPSCs, scale bar indicates 40 pA and 10 ms. A3 , Effect of experimental amiloride conditions on charge transfer. Series B, NHE block was achieved by substituting extracellular NaCl with LiCl. B1, Raw trace sequence from one CGC illustrates the effects of LiCl replacement, increased buffering capacity and a combination of the two conditions. B2 , averaged traces from one CGC under the conditions specified. B3 , effects of lithium on charge transfer. n=9 to 19 neurons in amiloride experiments, n=3 in lithium experiments. ★ p

    Techniques Used: Inhibition, Blocking Assay, Sequencing, Mass Spectrometry

    16) Product Images from "Acid-sensing ion channel 1a contributes to the effect of extracellular acidosis on NLRP1 inflammasome activation in cortical neurons"

    Article Title: Acid-sensing ion channel 1a contributes to the effect of extracellular acidosis on NLRP1 inflammasome activation in cortical neurons

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-015-0465-7

    ASICs mediate the increase of BK currents and the decrease of [K + ] i induced by extracellular acidosis. a Representative traces and statistical results showing amiloride did not influence BK currents at neutral pH. b Representative traces and statistical results showing amiloride inhibited acidosis-induced increase of BK currents. c Representative traces and statistical results showing PcTX1 did not influence BK currents at neutral pH. d Representative traces and statistical results showing PcTX1 inhibited acidosis-induced increase of BK currents. e , f Representative traces and statistical results showing acidosis failed to increase BK currents after pretreatment with PcTX1. g PcTX1 attenuated acidosis-induced decrease in [K + ] i . Data are expressed as means ± SEM. n = 6, # p
    Figure Legend Snippet: ASICs mediate the increase of BK currents and the decrease of [K + ] i induced by extracellular acidosis. a Representative traces and statistical results showing amiloride did not influence BK currents at neutral pH. b Representative traces and statistical results showing amiloride inhibited acidosis-induced increase of BK currents. c Representative traces and statistical results showing PcTX1 did not influence BK currents at neutral pH. d Representative traces and statistical results showing PcTX1 inhibited acidosis-induced increase of BK currents. e , f Representative traces and statistical results showing acidosis failed to increase BK currents after pretreatment with PcTX1. g PcTX1 attenuated acidosis-induced decrease in [K + ] i . Data are expressed as means ± SEM. n = 6, # p

    Techniques Used:

    17) Product Images from "pH-activated, mitochondria-targeted, and redox-responsive delivery of paclitaxel nanomicelles to overcome drug resistance and suppress metastasis in lung cancer"

    Article Title: pH-activated, mitochondria-targeted, and redox-responsive delivery of paclitaxel nanomicelles to overcome drug resistance and suppress metastasis in lung cancer

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-021-00895-4

    Cellular uptake and lysosomal escape of DA-P-SS-T/PTX nanomicelles. A CLSM images of A549 cells pretreated with chlorpromazine, genistein, and amiloride and then treated with DA-P-SS-T/C6 nanomicelles. The nuclei were stained by DAPI (blue). * P
    Figure Legend Snippet: Cellular uptake and lysosomal escape of DA-P-SS-T/PTX nanomicelles. A CLSM images of A549 cells pretreated with chlorpromazine, genistein, and amiloride and then treated with DA-P-SS-T/C6 nanomicelles. The nuclei were stained by DAPI (blue). * P

    Techniques Used: Confocal Laser Scanning Microscopy, Staining

    18) Product Images from "Regulation of intracellular pH during H+-coupled oligopeptide absorption in enterocytes from guinea-pig ileum"

    Article Title: Regulation of intracellular pH during H+-coupled oligopeptide absorption in enterocytes from guinea-pig ileum

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.1998.573bh.x

    Dose-response relationship for the inhibition of pH i recovery from Gly-Gly-imposed acid loading by amiloride in the CO 2 /HCO 3 − -buffered solution An acid load was imposed on cells by pulsed perfusion with 10 mM Gly-Gly, and the initial rate of pH i recovery was determined in the presence of various concentrations of amiloride. n = 4–6 for each concentration of amiloride. The curve was determined by fitting the data to the Michaelis-Menten equation.
    Figure Legend Snippet: Dose-response relationship for the inhibition of pH i recovery from Gly-Gly-imposed acid loading by amiloride in the CO 2 /HCO 3 − -buffered solution An acid load was imposed on cells by pulsed perfusion with 10 mM Gly-Gly, and the initial rate of pH i recovery was determined in the presence of various concentrations of amiloride. n = 4–6 for each concentration of amiloride. The curve was determined by fitting the data to the Michaelis-Menten equation.

    Techniques Used: Inhibition, Concentration Assay

    Effect of amiloride on pH i recovery from acid loading imposed by Gly-Gly A , representative trace in the CO 2 /HCO 3 − -buffered solution. Gly-Gly was added to the perfusate at a concentration of 10 mM by replacing equimolar mannitol. Dashed lines show the initial rates of pH i recovery. B , representative trace in the Hepes-buffered solution. C , effect of 0.3 mM amiloride on H + efflux after removing Gly-Gly in the presence and absence of CO 2 and HCO 3 − . The control value is the mean value of H + efflux from the first measurement without an inhibitor and the final measurement once the inhibitor had been removed. Each H + -equivalent efflux was estimated from the initial rate of pH i recovery and buffer capacity. n = 4 for the Hepes- and n = 8 for the CO 2 /HCO 3 − -buffered solution. * P
    Figure Legend Snippet: Effect of amiloride on pH i recovery from acid loading imposed by Gly-Gly A , representative trace in the CO 2 /HCO 3 − -buffered solution. Gly-Gly was added to the perfusate at a concentration of 10 mM by replacing equimolar mannitol. Dashed lines show the initial rates of pH i recovery. B , representative trace in the Hepes-buffered solution. C , effect of 0.3 mM amiloride on H + efflux after removing Gly-Gly in the presence and absence of CO 2 and HCO 3 − . The control value is the mean value of H + efflux from the first measurement without an inhibitor and the final measurement once the inhibitor had been removed. Each H + -equivalent efflux was estimated from the initial rate of pH i recovery and buffer capacity. n = 4 for the Hepes- and n = 8 for the CO 2 /HCO 3 − -buffered solution. * P

    Techniques Used: Concentration Assay

    Na + -dependent pH i recovery from acid loading imposed by pulsed NH 4 Cl and the effect of amiloride on pH i recovery in the Hepes-buffered solution The villus tip was acid-loaded 3 times by being exposed to 40 mM NH 4 Cl for 90 s. Dashed lines indicate the initial rate of pH i recovery. A representative trace from 6 experiments is presented.
    Figure Legend Snippet: Na + -dependent pH i recovery from acid loading imposed by pulsed NH 4 Cl and the effect of amiloride on pH i recovery in the Hepes-buffered solution The villus tip was acid-loaded 3 times by being exposed to 40 mM NH 4 Cl for 90 s. Dashed lines indicate the initial rate of pH i recovery. A representative trace from 6 experiments is presented.

    Techniques Used:

    19) Product Images from "Osteoclast-independent bone resorption by fibroblast-like cells"

    Article Title: Osteoclast-independent bone resorption by fibroblast-like cells

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar752

    Release of acidic components by prosthesis loosening fibroblasts (PLFs). (a) and (b) Cytosensor measurement of PLFs stimulated with tumour necrosis factor (TNF)-α (1, 10 and 300 ng/ml) revealed the release of acidic components, with maximal pericellular acidification (rmax/req) of 15% at 10 ng/ml TNF-α (A). Recording of the time curve with 10 ng/ml TNF-α showed a maximal acidification 15 min after the influx of TNF-α was started (b). (c) Incubation of PLFs with the ATPase inhibitors amiloride and bafilomycin A 1 at different concentrations decreased the pericellular acidification by a maximum of 32% with amiloride and 11% with bafilomycin A 1 . (d) Inhibition of H + release by amiloride was recorded after 3 min and remained stable for the total measuring time of 45 min. Pericellular pH returned to the initial values shortly after the perfusion of amiloride was terminated. (e) The specific v-ATPase inhibitor bafilomycin A 1 at concentrations of 10 -6 mol/l showed a clear effect shortly after its addition, and the H + secretion remained low even after discontinuation of bafilomycin A 1 infusion.
    Figure Legend Snippet: Release of acidic components by prosthesis loosening fibroblasts (PLFs). (a) and (b) Cytosensor measurement of PLFs stimulated with tumour necrosis factor (TNF)-α (1, 10 and 300 ng/ml) revealed the release of acidic components, with maximal pericellular acidification (rmax/req) of 15% at 10 ng/ml TNF-α (A). Recording of the time curve with 10 ng/ml TNF-α showed a maximal acidification 15 min after the influx of TNF-α was started (b). (c) Incubation of PLFs with the ATPase inhibitors amiloride and bafilomycin A 1 at different concentrations decreased the pericellular acidification by a maximum of 32% with amiloride and 11% with bafilomycin A 1 . (d) Inhibition of H + release by amiloride was recorded after 3 min and remained stable for the total measuring time of 45 min. Pericellular pH returned to the initial values shortly after the perfusion of amiloride was terminated. (e) The specific v-ATPase inhibitor bafilomycin A 1 at concentrations of 10 -6 mol/l showed a clear effect shortly after its addition, and the H + secretion remained low even after discontinuation of bafilomycin A 1 infusion.

    Techniques Used: Incubation, Inhibition

    20) Product Images from "The Influence of Nano-Carrier Architecture on In Vitro siRNA Delivery Performance and In Vivo Biodistribution: Polyplexes vs. Micelleplexes"

    Article Title: The Influence of Nano-Carrier Architecture on In Vitro siRNA Delivery Performance and In Vivo Biodistribution: Polyplexes vs. Micelleplexes

    Journal: ACS Nano

    doi: 10.1021/nn102540y

    Flow cytometry analysis of AlexaFluor647 uptake in HeLa cells facilitated by various delivery vehicles in the presence or absence of endocytosis inhibitors. Amiloride, amantadine, nystatin, methyl β-cyclodextrin, folic acid, and PDMP hydrochloride
    Figure Legend Snippet: Flow cytometry analysis of AlexaFluor647 uptake in HeLa cells facilitated by various delivery vehicles in the presence or absence of endocytosis inhibitors. Amiloride, amantadine, nystatin, methyl β-cyclodextrin, folic acid, and PDMP hydrochloride

    Techniques Used: Flow Cytometry, Cytometry

    21) Product Images from "Cerebral influx of Na+ and Cl− as the osmotherapy-mediated rebound response in rats"

    Article Title: Cerebral influx of Na+ and Cl− as the osmotherapy-mediated rebound response in rats

    Journal: Fluids and Barriers of the CNS

    doi: 10.1186/s12987-018-0111-8

    Inhibitors of ion-transporting mechanisms at the blood-side membranes do not affect water loss and electrolyte gain. a The arterial blood pressure was measured before and until 1 h after i.v. treatment with vehicle or inhibitors (10 mg/kg bumetanide, 6 mg/kg amiloride, and 20 mg/kg methazolamide). Values are given as the percentage of arterial blood pressure from the last control measurement (corresponding to 30 s before i.v. injection). The arterial blood pressure did not differ significantly from control measurements after 1 h (p > 0.90). The end arterial blood pressure was unchanged following inhibitor delivery, n = 3 of each, p > 0.90. b The brain water content was unaffected by i.v. inhibitor application in control rats [in (ml/g dry weight): vehicle: 3.79 ± 0.01 vs. inhibitors: 3.76 ± 0.01] and in rats subjected to NaCl-mediated osmotherapy (vehicle: 3.46 ± 0.01 vs. inhibitors: 3.45 ± 0.02), n = 7–9. Inset: Brain water content in osmotherapy-treated rats exposed to triple doses of vehicle (3.38 ± 0.02) or inhibitors (3.38 ± 0.02), n = 4 of each. c The brain Na + content (in mmol/kg dry weight) in control rats (vehicle: 197 ± 1 vs. inhibitors: 194 ± 1) and in rats exposed to osmotherapy (vehicle: 227 ± 2 vs. inhibitors: 224 ± 3), n = 7–9. d The brain Cl − content (in mmol/kg dry weight) in control rats (vehicle: 132 ± 3 vs. inhibitors: 131 ± 4) and in rats exposed to osmotherapy (vehicle: 173 ± 3 vs. inhibitors: 170 ± 4), n = 7–9. Vehicle values from control and osmotherapy-treated rats are from Fig. 2 a–c and included for comparison. Statistically significant differences were determined by a two-way ANOVA with Tukey’s multiple comparisons post hoc test, except for values in the inset of b , which were analyzed using a two-tailed un-paired Student’s t-test. ns not significant
    Figure Legend Snippet: Inhibitors of ion-transporting mechanisms at the blood-side membranes do not affect water loss and electrolyte gain. a The arterial blood pressure was measured before and until 1 h after i.v. treatment with vehicle or inhibitors (10 mg/kg bumetanide, 6 mg/kg amiloride, and 20 mg/kg methazolamide). Values are given as the percentage of arterial blood pressure from the last control measurement (corresponding to 30 s before i.v. injection). The arterial blood pressure did not differ significantly from control measurements after 1 h (p > 0.90). The end arterial blood pressure was unchanged following inhibitor delivery, n = 3 of each, p > 0.90. b The brain water content was unaffected by i.v. inhibitor application in control rats [in (ml/g dry weight): vehicle: 3.79 ± 0.01 vs. inhibitors: 3.76 ± 0.01] and in rats subjected to NaCl-mediated osmotherapy (vehicle: 3.46 ± 0.01 vs. inhibitors: 3.45 ± 0.02), n = 7–9. Inset: Brain water content in osmotherapy-treated rats exposed to triple doses of vehicle (3.38 ± 0.02) or inhibitors (3.38 ± 0.02), n = 4 of each. c The brain Na + content (in mmol/kg dry weight) in control rats (vehicle: 197 ± 1 vs. inhibitors: 194 ± 1) and in rats exposed to osmotherapy (vehicle: 227 ± 2 vs. inhibitors: 224 ± 3), n = 7–9. d The brain Cl − content (in mmol/kg dry weight) in control rats (vehicle: 132 ± 3 vs. inhibitors: 131 ± 4) and in rats exposed to osmotherapy (vehicle: 173 ± 3 vs. inhibitors: 170 ± 4), n = 7–9. Vehicle values from control and osmotherapy-treated rats are from Fig. 2 a–c and included for comparison. Statistically significant differences were determined by a two-way ANOVA with Tukey’s multiple comparisons post hoc test, except for values in the inset of b , which were analyzed using a two-tailed un-paired Student’s t-test. ns not significant

    Techniques Used: Injection, Two Tailed Test

    22) Product Images from "Synergistic effects of ion transporter and MAP kinase pathway inhibitors in melanoma"

    Article Title: Synergistic effects of ion transporter and MAP kinase pathway inhibitors in melanoma

    Journal: Nature Communications

    doi: 10.1038/ncomms12336

    Digitoxin and MEK inhibitor additively or synergistically reduced intracellular pH in melanoma cells by synergistically inhibiting the NHE Na + /H + exchanger. ( a ) Intracellular pH in live melanoma cells acutely dissociated from tumours treated for 4 days in vivo (each melanoma was tested in two independent experiments). ( b ) Intracellular pH of human and mouse cells in the blood of NSG mice xenografted with hUCB cells and treated with digitoxin plus MEK inhibitor for 10 days. ( c ) Intracellular pH of immortalized melanocytes (hiMEL) and M214 melanoma cells grown subcutaneously in gelatin sponges in NSG mice that were treated with digitoxin plus MEK inhibitor for four days ( n =2 independent experiments). ( d ) NHE activity in dissociated tumour cells isolated from mice treated 4 days in vivo with digitoxin and/or MEK inhibitor was measured based on their rate of recovery from sodium propionate-induced acute intracellular acidification (each melanoma was tested in 2–3 independent experiments). ( e , f ) Intracellular pH in dissociated cells ( e ) and activated caspase-3 + cells in sections ( f ) from melanomas (M481 and M214) obtained from mice treated for 4 days with amiloride (NHE inhibitor) or digitoxin plus MEK inhibitor ( n =4–9 mice per treatment; each melanoma was tested in 1–2 independent experiments). In these experiments, an earlier passage of M214 cells was used, with a higher intracellular pH, compared with the M214 cells shown in a and g . ( g , h ) Intracellular pH in dissociated tumour cells ( g ) and activated caspase-3 + cells in sections ( h ) of melanoma xenografts (M481 and M214) expressing vector (tRFP control) or NHE1 and treated 4 days with digitoxin plus MEK inhibitor (each melanoma was tested in an independent experiment). ( i ) Intracellular pH in dissociated tumour cells from M481 xenografts that were uninfected, expressing vector (tRFP control) or BCL2 and treated 4 days with digitoxin plus MEK inhibitor. Statistical significance was assessed by one-way analysis of variance followed by Dunnett's multiple comparisons test. All data represent mean±s.d. In each panel the number of mice per treatment is written on the bars.
    Figure Legend Snippet: Digitoxin and MEK inhibitor additively or synergistically reduced intracellular pH in melanoma cells by synergistically inhibiting the NHE Na + /H + exchanger. ( a ) Intracellular pH in live melanoma cells acutely dissociated from tumours treated for 4 days in vivo (each melanoma was tested in two independent experiments). ( b ) Intracellular pH of human and mouse cells in the blood of NSG mice xenografted with hUCB cells and treated with digitoxin plus MEK inhibitor for 10 days. ( c ) Intracellular pH of immortalized melanocytes (hiMEL) and M214 melanoma cells grown subcutaneously in gelatin sponges in NSG mice that were treated with digitoxin plus MEK inhibitor for four days ( n =2 independent experiments). ( d ) NHE activity in dissociated tumour cells isolated from mice treated 4 days in vivo with digitoxin and/or MEK inhibitor was measured based on their rate of recovery from sodium propionate-induced acute intracellular acidification (each melanoma was tested in 2–3 independent experiments). ( e , f ) Intracellular pH in dissociated cells ( e ) and activated caspase-3 + cells in sections ( f ) from melanomas (M481 and M214) obtained from mice treated for 4 days with amiloride (NHE inhibitor) or digitoxin plus MEK inhibitor ( n =4–9 mice per treatment; each melanoma was tested in 1–2 independent experiments). In these experiments, an earlier passage of M214 cells was used, with a higher intracellular pH, compared with the M214 cells shown in a and g . ( g , h ) Intracellular pH in dissociated tumour cells ( g ) and activated caspase-3 + cells in sections ( h ) of melanoma xenografts (M481 and M214) expressing vector (tRFP control) or NHE1 and treated 4 days with digitoxin plus MEK inhibitor (each melanoma was tested in an independent experiment). ( i ) Intracellular pH in dissociated tumour cells from M481 xenografts that were uninfected, expressing vector (tRFP control) or BCL2 and treated 4 days with digitoxin plus MEK inhibitor. Statistical significance was assessed by one-way analysis of variance followed by Dunnett's multiple comparisons test. All data represent mean±s.d. In each panel the number of mice per treatment is written on the bars.

    Techniques Used: In Vivo, Mouse Assay, Activity Assay, Isolation, Expressing, Plasmid Preparation

    23) Product Images from "Regulation of ENaC-mediated alveolar fluid clearance by insulin via PI3K/Akt pathway in LPS-induced acute lung injury"

    Article Title: Regulation of ENaC-mediated alveolar fluid clearance by insulin via PI3K/Akt pathway in LPS-induced acute lung injury

    Journal: Respiratory Research

    doi: 10.1186/1465-9921-13-29

    Effect of exogenous insulin on pulmonary edema and alveolar fluid clearance in LPS-induced actue lung injury (n = 6 per group) . (A) Total lung water content at 0, 2, 4, 8 hours after LPS or insulin treatment. (B) Alveolar fluid clearance at 0, 2, 4, 8 hours after LPS or insulin treatment. (C) Total lung water content at 8 hours after LPS-induced actue lung injury or saline treatment (D) Alveolar fluid clearance at 8 hours after LPS-induced actue lung injury or saline treatment. Albumin solution containing amiloride (5 × 10 -4 M) were injected into the alveolar spaces. Data are presented as mean ± S.E.M.Δ P
    Figure Legend Snippet: Effect of exogenous insulin on pulmonary edema and alveolar fluid clearance in LPS-induced actue lung injury (n = 6 per group) . (A) Total lung water content at 0, 2, 4, 8 hours after LPS or insulin treatment. (B) Alveolar fluid clearance at 0, 2, 4, 8 hours after LPS or insulin treatment. (C) Total lung water content at 8 hours after LPS-induced actue lung injury or saline treatment (D) Alveolar fluid clearance at 8 hours after LPS-induced actue lung injury or saline treatment. Albumin solution containing amiloride (5 × 10 -4 M) were injected into the alveolar spaces. Data are presented as mean ± S.E.M.Δ P

    Techniques Used: Injection

    24) Product Images from "Deleterious Effects of Plasminogen Activators in Neonatal Cerebral Hypoxia-Ischemia"

    Article Title: Deleterious Effects of Plasminogen Activators in Neonatal Cerebral Hypoxia-Ischemia

    Journal: The American Journal of Pathology

    doi: 10.2353/ajpath.2008.070979

    Induction of the plasminogen system correlates with HI-damaged brain regions. A: In situ gel zymogram using unfixed brain sections showed increased PA activity on the carotid-occluded side (R) at 4 hours after HI. The PA activity is concentrated in the dorsal cerebral cortex (Ctx) and hippocampus (Hip) than in the basal thalamus (Th). B: Omission of plasminogen in the overlay abolished the in situ proteolysis activity. C and D: Inclusion of a synthetic tPA inhibitor tPA-STOP (17 μmol/L) or a uPA inhibitor amiloride (0.1 mmol/L) reduced the extent of proteolysis. Images shown are representative results from at least three sets of samples. E: Representative examples of Nissl stain indicating damage in the posterior part of the forebrain at 7 days of recovery after HI. Note the typical pathology of obliteration of the hippocampus and unilateral dilation of the cerebral ventricle ( arrows ), as well as cystic degeneration of the dorsolateral cerebral cortex ( arrowheads ).
    Figure Legend Snippet: Induction of the plasminogen system correlates with HI-damaged brain regions. A: In situ gel zymogram using unfixed brain sections showed increased PA activity on the carotid-occluded side (R) at 4 hours after HI. The PA activity is concentrated in the dorsal cerebral cortex (Ctx) and hippocampus (Hip) than in the basal thalamus (Th). B: Omission of plasminogen in the overlay abolished the in situ proteolysis activity. C and D: Inclusion of a synthetic tPA inhibitor tPA-STOP (17 μmol/L) or a uPA inhibitor amiloride (0.1 mmol/L) reduced the extent of proteolysis. Images shown are representative results from at least three sets of samples. E: Representative examples of Nissl stain indicating damage in the posterior part of the forebrain at 7 days of recovery after HI. Note the typical pathology of obliteration of the hippocampus and unilateral dilation of the cerebral ventricle ( arrows ), as well as cystic degeneration of the dorsolateral cerebral cortex ( arrowheads ).

    Techniques Used: In Situ, Activity Assay, Staining

    25) Product Images from "Insulin activates epithelial sodium channel (ENaC) via phosphoinositide 3-kinase in mammalian taste receptor cells"

    Article Title: Insulin activates epithelial sodium channel (ENaC) via phosphoinositide 3-kinase in mammalian taste receptor cells

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00318.2010

    The importance of serum- and glucocorticoid-regulated kinase (SGK) in insulin/ENaC mediated responses in taste cells. A : functional Na + imaging with SBFI from SGK +/+ wild-type mice in the presence of insulin (20 nM) and, subsequently, amiloride (30 μM).
    Figure Legend Snippet: The importance of serum- and glucocorticoid-regulated kinase (SGK) in insulin/ENaC mediated responses in taste cells. A : functional Na + imaging with SBFI from SGK +/+ wild-type mice in the presence of insulin (20 nM) and, subsequently, amiloride (30 μM).

    Techniques Used: Functional Assay, Imaging, Mouse Assay

    Effects of insulin on amiloride-sensitive cells. A : steady-state current in a mouse taste cell during whole cell patch-clamp recording. Shown is a continuous recording from a fungiform taste cell during the application of insulin (10 nM), insulin with
    Figure Legend Snippet: Effects of insulin on amiloride-sensitive cells. A : steady-state current in a mouse taste cell during whole cell patch-clamp recording. Shown is a continuous recording from a fungiform taste cell during the application of insulin (10 nM), insulin with

    Techniques Used: Patch Clamp

    Insulin-induced effects on amiloride-sensitive cells were abolished by both wortmannin and LY294002. A : insulin enhancement on Na + influx was recorded with functional Na + imaging in the absence and presence of wortmannin (1 μM) treatment for 1
    Figure Legend Snippet: Insulin-induced effects on amiloride-sensitive cells were abolished by both wortmannin and LY294002. A : insulin enhancement on Na + influx was recorded with functional Na + imaging in the absence and presence of wortmannin (1 μM) treatment for 1

    Techniques Used: Functional Assay, Imaging

    26) Product Images from "The Effects of Amiloride and Age on Oxygen Consumption Coupled to Electrogenic Sodium Transport in the Human Sigmoid Colon"

    Article Title: The Effects of Amiloride and Age on Oxygen Consumption Coupled to Electrogenic Sodium Transport in the Human Sigmoid Colon

    Journal: Saudi Journal of Gastroenterology : Official Journal of the Saudi Gastroenterology Association

    doi: 10.4103/1319-3767.164190

    Regression analysis of short-circuit current (μA/cm 2 , upper panel) and oxygen consumption rate (micromol/h/cm 2 , lower panel) versus age under baseline conditions (circles) and after addition of amiloride (squares). The 95% confidence intervals are shown in trace lines. No regression line was significantly different from zero
    Figure Legend Snippet: Regression analysis of short-circuit current (μA/cm 2 , upper panel) and oxygen consumption rate (micromol/h/cm 2 , lower panel) versus age under baseline conditions (circles) and after addition of amiloride (squares). The 95% confidence intervals are shown in trace lines. No regression line was significantly different from zero

    Techniques Used:

    27) Product Images from "Evaluating the Functionality of Conjunctiva Using a Rabbit Dry Eye Model"

    Article Title: Evaluating the Functionality of Conjunctiva Using a Rabbit Dry Eye Model

    Journal: Journal of Ophthalmology

    doi: 10.1155/2016/3964642

    Effect of epithelial sodium channel blockers on rabbit DE model. The application of epithelial sodium channel blockers, benzamil (a) and amiloride (b), did not significantly increase the tear quantity in our rabbit DE model.
    Figure Legend Snippet: Effect of epithelial sodium channel blockers on rabbit DE model. The application of epithelial sodium channel blockers, benzamil (a) and amiloride (b), did not significantly increase the tear quantity in our rabbit DE model.

    Techniques Used:

    Potential difference recordings of the rabbit eyes after surgery compared to rabbit eyes without surgery. The potential differences were recorded for the rabbit eyes 5 months AE ( n = 4) and control eyes ( n = 4). The perfusion channel was switched from PBS to amiloride at 13 seconds, with 2-3 seconds required for the new solution to reach the ocular surface. Data are presented as mean Standard Error Method (SEM).
    Figure Legend Snippet: Potential difference recordings of the rabbit eyes after surgery compared to rabbit eyes without surgery. The potential differences were recorded for the rabbit eyes 5 months AE ( n = 4) and control eyes ( n = 4). The perfusion channel was switched from PBS to amiloride at 13 seconds, with 2-3 seconds required for the new solution to reach the ocular surface. Data are presented as mean Standard Error Method (SEM).

    Techniques Used:

    28) Product Images from "Catabolic pathways regulated by mTORC1 are pivotal for survival and growth of cancer cells expressing mutant Ras"

    Article Title: Catabolic pathways regulated by mTORC1 are pivotal for survival and growth of cancer cells expressing mutant Ras

    Journal: Oncotarget

    doi:

    Autophagy and macropinocytosis inhibition sensitize oncogenic Ras cells in combinatorial treatment with mTOR inhibitor A. MIA PaCa-2 (KRas mutant) cells were treated with the indicated concentrations of CQ, amiloride, or rapamycin alone, or two of each in combination, for 72 hours. Cellular growth was assessed using an MTT assay. B. MIA PaCa-2 cells expressing Atg7 shRNA and control shRNA were treated with the indicated concentrations of rapamycin for 48 h and cell growth was assessed using an MTT assay. Cell growth with 5 μM rapamycin treatment was monitored by cell counting in MIA PaCa-2 expressing both Atg7 shRNA and control shRNA. Relative growth is presented after normalization by number of cells under the each condition without rapamycin treatment. C. Concomitant treatment with rapamycin and CQ or rapamycin and amiloride resulted in the inhibition of tumor growth in a xenograft mouse model. Subcutaneous MIA PaCa-2-driven tumors were established in 6-week old male mice. Rapamycin (1 mg kg −1 per day), CQ (20 mg kg −1 per day), or amiloride (10 mg kg −1 per day) alone, or two of each in combination, were administered daily via intraperitoneal injection. Tumor growth was assessed once tumor volume reached 150 mm 3 . Data are shown as the mean of five mice in each group +/−SEM. * p
    Figure Legend Snippet: Autophagy and macropinocytosis inhibition sensitize oncogenic Ras cells in combinatorial treatment with mTOR inhibitor A. MIA PaCa-2 (KRas mutant) cells were treated with the indicated concentrations of CQ, amiloride, or rapamycin alone, or two of each in combination, for 72 hours. Cellular growth was assessed using an MTT assay. B. MIA PaCa-2 cells expressing Atg7 shRNA and control shRNA were treated with the indicated concentrations of rapamycin for 48 h and cell growth was assessed using an MTT assay. Cell growth with 5 μM rapamycin treatment was monitored by cell counting in MIA PaCa-2 expressing both Atg7 shRNA and control shRNA. Relative growth is presented after normalization by number of cells under the each condition without rapamycin treatment. C. Concomitant treatment with rapamycin and CQ or rapamycin and amiloride resulted in the inhibition of tumor growth in a xenograft mouse model. Subcutaneous MIA PaCa-2-driven tumors were established in 6-week old male mice. Rapamycin (1 mg kg −1 per day), CQ (20 mg kg −1 per day), or amiloride (10 mg kg −1 per day) alone, or two of each in combination, were administered daily via intraperitoneal injection. Tumor growth was assessed once tumor volume reached 150 mm 3 . Data are shown as the mean of five mice in each group +/−SEM. * p

    Techniques Used: Inhibition, Mutagenesis, MTT Assay, Expressing, shRNA, Cell Counting, Mouse Assay, Injection

    29) Product Images from "Amiloride and guggulsterone suppression of esophageal cancer cell growth in vitro and in nude mouse xenografts"

    Article Title: Amiloride and guggulsterone suppression of esophageal cancer cell growth in vitro and in nude mouse xenografts

    Journal: Frontiers in biology

    doi: 10.1007/s11515-014-1289-z

    Effect of NHE-1 or FXR inhibition on regulation of esophageal cancer cell viability and apoptosis. A, MTT assay. Esophageal cancer cells TE-3, TE-12, SKGT-4, and SKGT-5 were grown and treated with 200 μM amiloride, 25 μM guggulsterone,
    Figure Legend Snippet: Effect of NHE-1 or FXR inhibition on regulation of esophageal cancer cell viability and apoptosis. A, MTT assay. Esophageal cancer cells TE-3, TE-12, SKGT-4, and SKGT-5 were grown and treated with 200 μM amiloride, 25 μM guggulsterone,

    Techniques Used: Inhibition, MTT Assay

    Effect of NHE-1 and FXR inhibition in suppression of growth of nude mouse xenografts. The mice were treated with 5 mg/kg amiloride, 50 mg/kg guggulsterone, or a combination of both (as the same individual dose) for 2 days before they were subcutaneously
    Figure Legend Snippet: Effect of NHE-1 and FXR inhibition in suppression of growth of nude mouse xenografts. The mice were treated with 5 mg/kg amiloride, 50 mg/kg guggulsterone, or a combination of both (as the same individual dose) for 2 days before they were subcutaneously

    Techniques Used: Inhibition, Mouse Assay

    30) Product Images from "A Switch in the Mechanism of Hypertension in the Syndrome of Apparent Mineralocorticoid Excess"

    Article Title: A Switch in the Mechanism of Hypertension in the Syndrome of Apparent Mineralocorticoid Excess

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2007040401

    (A) Fractional sodium excretion in 80-d-old (7/5) and 120-d-old (7/7) 11βHSD2 −/− mice (▪) and age-matched C57BL/6J mice (□) (B) The absolute effect of amiloride on sodium excretion (Δ amiloride). (C) The
    Figure Legend Snippet: (A) Fractional sodium excretion in 80-d-old (7/5) and 120-d-old (7/7) 11βHSD2 −/− mice (▪) and age-matched C57BL/6J mice (□) (B) The absolute effect of amiloride on sodium excretion (Δ amiloride). (C) The

    Techniques Used: Mouse Assay

    31) Product Images from "Male Sex is Associated with a Reduced Alveolar Epithelial Sodium Transport"

    Article Title: Male Sex is Associated with a Reduced Alveolar Epithelial Sodium Transport

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0136178

    Female FDLE cells display an increased Na + transport. (A) I base was the I SC after mounting the monolayers in the Ussing chambers, the amiloride-sensitive (I amil ) current represents the current reduction induced by amiloride (10 μM). I base and I amil , but not the amiloride-insensitive I SC , were significantly higher in FDLE cells of female origin (n = 45/56; mean + SEM; ***p
    Figure Legend Snippet: Female FDLE cells display an increased Na + transport. (A) I base was the I SC after mounting the monolayers in the Ussing chambers, the amiloride-sensitive (I amil ) current represents the current reduction induced by amiloride (10 μM). I base and I amil , but not the amiloride-insensitive I SC , were significantly higher in FDLE cells of female origin (n = 45/56; mean + SEM; ***p

    Techniques Used:

    Female FDLE monolayers show a higher ENaC and Na,K-ATPase activity in permeabilized membrane measurements. After the I SC reached a plateau (I base ), amphotericin B was used to permeabilize either the basolateral (100 μM) or the apical (10 μM) membrane. At the maximal current increase either amiloride (10 μM) or ouabain (1 mM) were applied and amil max (A) or ouab max (C) determined. I base , amil max and ouab max were significantly higher in FDLE cells of female origin (A: n = 35/32; C: n = 53/51; mean + SEM, *p
    Figure Legend Snippet: Female FDLE monolayers show a higher ENaC and Na,K-ATPase activity in permeabilized membrane measurements. After the I SC reached a plateau (I base ), amphotericin B was used to permeabilize either the basolateral (100 μM) or the apical (10 μM) membrane. At the maximal current increase either amiloride (10 μM) or ouabain (1 mM) were applied and amil max (A) or ouab max (C) determined. I base , amil max and ouab max were significantly higher in FDLE cells of female origin (A: n = 35/32; C: n = 53/51; mean + SEM, *p

    Techniques Used: Activity Assay

    32) Product Images from "Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells"

    Article Title: Impact of Na+ permeation on collective migration of pulmonary arterial endothelial cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0250095

    Inhibition of Na + permeation through ENaC and NCX channels slowed PAEC scratch wound closure. (A) Brightfield images of scratched PAEC monolayers at 0, 6, and 24-hours in vehicle control, 90 μM amiloride, and 10 μM benzamil conditions. Benzamil treatment caused accumulation of floating particles over 24 hours. Studies were all conducted in Na + -containing media. Scatter plots of wound closure distance in (B) 0–6, (C) 6–24, and (D) 0–24 hours. Benzamil was more potent than amiloride to slow PAEC monolayer wound closure. Data were reported as mean ± SEM. (E) Magnified view of the marginal regions of the control, amiloride-treated, and benzamil-treated cells after 24 hours of migration. An arrowhead points to a floating particle. Statistical significance was assessed using one-way ANOVA with Bonferroni post hoc test (ns—not significant, *— P
    Figure Legend Snippet: Inhibition of Na + permeation through ENaC and NCX channels slowed PAEC scratch wound closure. (A) Brightfield images of scratched PAEC monolayers at 0, 6, and 24-hours in vehicle control, 90 μM amiloride, and 10 μM benzamil conditions. Benzamil treatment caused accumulation of floating particles over 24 hours. Studies were all conducted in Na + -containing media. Scatter plots of wound closure distance in (B) 0–6, (C) 6–24, and (D) 0–24 hours. Benzamil was more potent than amiloride to slow PAEC monolayer wound closure. Data were reported as mean ± SEM. (E) Magnified view of the marginal regions of the control, amiloride-treated, and benzamil-treated cells after 24 hours of migration. An arrowhead points to a floating particle. Statistical significance was assessed using one-way ANOVA with Bonferroni post hoc test (ns—not significant, *— P

    Techniques Used: Inhibition, Migration

    33) Product Images from "Acid-sensing ion channels contribute to the metaboreceptor component of the exercise pressor reflex"

    Article Title: Acid-sensing ion channels contribute to the metaboreceptor component of the exercise pressor reflex

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    doi: 10.1152/ajpheart.00328.2009

    Effects of amiloride on pressor ( A ) and renal sympathetic nerve ( B ) responses to static contraction while the muscles were freely perfused and while the circulation was occluded both before and after amiloride. Bars represent means ± SE. * P
    Figure Legend Snippet: Effects of amiloride on pressor ( A ) and renal sympathetic nerve ( B ) responses to static contraction while the muscles were freely perfused and while the circulation was occluded both before and after amiloride. Bars represent means ± SE. * P

    Techniques Used:

    34) Product Images from "Endolymphatic Sodium Homeostasis by Extramacular Epithelium of the Saccule"

    Article Title: Endolymphatic Sodium Homeostasis by Extramacular Epithelium of the Saccule

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3044-09.2009

    Effect of Na + transport inhibitors on current from saccular extramacular epithelium. A , Amiloride ( n = 5); bars show duration of perfusion at concentrations (molar) expressed as the negative log of the number in the bar. B , Benzamil ( n = 5). C , EIPA (
    Figure Legend Snippet: Effect of Na + transport inhibitors on current from saccular extramacular epithelium. A , Amiloride ( n = 5); bars show duration of perfusion at concentrations (molar) expressed as the negative log of the number in the bar. B , Benzamil ( n = 5). C , EIPA (

    Techniques Used:

    35) Product Images from "ADENOSINE ACTIVATES A2b RECEPTORS AND ENHANCES CHLORIDE SECRETION IN KIDNEY INNER MEDULLARY COLLECTING DUCT CELLS"

    Article Title: ADENOSINE ACTIVATES A2b RECEPTORS AND ENHANCES CHLORIDE SECRETION IN KIDNEY INNER MEDULLARY COLLECTING DUCT CELLS

    Journal: Hypertension

    doi: 10.1161/HYPERTENSIONAHA.109.143404

    A. Representative I sc trace showing actions of the specific A2a receptor antagonist SCH442416 and the specific A2b receptor antagonist PSB603 on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5
    Figure Legend Snippet: A. Representative I sc trace showing actions of the specific A2a receptor antagonist SCH442416 and the specific A2b receptor antagonist PSB603 on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5

    Techniques Used:

    A, B. Representative traces of I sc showing the effect of pre-incubation with vehicle control (H 2 O) or PKA-inhibitor H-89 (10 −5 mol/L) on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5
    Figure Legend Snippet: A, B. Representative traces of I sc showing the effect of pre-incubation with vehicle control (H 2 O) or PKA-inhibitor H-89 (10 −5 mol/L) on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5

    Techniques Used: Incubation

    A. Representative trace of I sc showing the effects of CFTR inhibitor-172 and DIDS on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5 mol/L adenosine (apical addition); CI, 10 −5 mol/L
    Figure Legend Snippet: A. Representative trace of I sc showing the effects of CFTR inhibitor-172 and DIDS on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10 −5 mol/L adenosine (apical addition); CI, 10 −5 mol/L

    Techniques Used:

    A. Representative I sc trace describing the action of the specific A1 receptor antagonist DPCPX and the non-selective A2 receptor antagonist ZM241385 on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10
    Figure Legend Snippet: A. Representative I sc trace describing the action of the specific A1 receptor antagonist DPCPX and the non-selective A2 receptor antagonist ZM241385 on adenosine-induced I sc in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, 10

    Techniques Used:

    A. Representative I sc trace of cumulative dose response to adenosine in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, adenosine concentrations in mol/L (M) as specified (addition to both sides); DPAC, 2 × 10 −3
    Figure Legend Snippet: A. Representative I sc trace of cumulative dose response to adenosine in mIMCD-K2 cells. Am, 10 −5 mol/L amiloride (apical addition); Ado, adenosine concentrations in mol/L (M) as specified (addition to both sides); DPAC, 2 × 10 −3

    Techniques Used:

    36) Product Images from "Using Drugs to Probe the Variability of Trans-Epithelial Airway Resistance"

    Article Title: Using Drugs to Probe the Variability of Trans-Epithelial Airway Resistance

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0149550

    Relationship between baseline voltage and response of TER after amiloride addition. Logarithm of baseline V plotted against ratio of TER upon amiloride addition in drug regime I, applying the initial baseline (A-I), or the rolling baseline approach (B-I). In drug regime II (A-II), grouping produced a significant difference between the two groups (LOW, n = 57; HIGH, n = 24). For both drug regimes, grouping was performed as in Figs 5 and 6 . Additional regression and statistical analysis data are shown in Table D, S1 File .
    Figure Legend Snippet: Relationship between baseline voltage and response of TER after amiloride addition. Logarithm of baseline V plotted against ratio of TER upon amiloride addition in drug regime I, applying the initial baseline (A-I), or the rolling baseline approach (B-I). In drug regime II (A-II), grouping produced a significant difference between the two groups (LOW, n = 57; HIGH, n = 24). For both drug regimes, grouping was performed as in Figs 5 and 6 . Additional regression and statistical analysis data are shown in Table D, S1 File .

    Techniques Used: Produced

    Drug induced effect on TER with respect to the initial baseline. TER values at baseline are plotted against values obtained after the cumulative addition of the different drugs. The hypotenuse of the shaded triangular area represents the line of identity, while magnifications are shown in the box inserts. (A-I) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of FSK (n = 44), (B-I) FSK+CFTR Inh172 (n = 44) and (C-I) FSK+CFTR Inh172 +AMI (n = 44); line of regression is shown as a dashed line with 99% CI (grey vertical bars). In D-I same data as in C-I showing two distinct populations of amiloride responders designated as LOW (closed grey circles-black dashed regression line +99% CI vertical bars, n = 31) and HIGH (open squares, black regression line +99% CI dotted line, n = 13). (A-II) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of amiloride; line of regression is shown as a dashed line with 99% CI (grey vertical bars). In B-II same data as in A-II showing two distinct populations designated as LOW (n = 57) and HIGH (n = 24) amiloride responders. Changes in TER with cumulative drug addition of (C-II) AMI+FSK (LOW n = 56, HIGH n = 24) and (D-II) AMI+FSK+CFTR Inh172 (LOW n = 56, HIGH n = 21). Additional regression and statistical analysis data are shown in Table B in S1 File .
    Figure Legend Snippet: Drug induced effect on TER with respect to the initial baseline. TER values at baseline are plotted against values obtained after the cumulative addition of the different drugs. The hypotenuse of the shaded triangular area represents the line of identity, while magnifications are shown in the box inserts. (A-I) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of FSK (n = 44), (B-I) FSK+CFTR Inh172 (n = 44) and (C-I) FSK+CFTR Inh172 +AMI (n = 44); line of regression is shown as a dashed line with 99% CI (grey vertical bars). In D-I same data as in C-I showing two distinct populations of amiloride responders designated as LOW (closed grey circles-black dashed regression line +99% CI vertical bars, n = 31) and HIGH (open squares, black regression line +99% CI dotted line, n = 13). (A-II) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of amiloride; line of regression is shown as a dashed line with 99% CI (grey vertical bars). In B-II same data as in A-II showing two distinct populations designated as LOW (n = 57) and HIGH (n = 24) amiloride responders. Changes in TER with cumulative drug addition of (C-II) AMI+FSK (LOW n = 56, HIGH n = 24) and (D-II) AMI+FSK+CFTR Inh172 (LOW n = 56, HIGH n = 21). Additional regression and statistical analysis data are shown in Table B in S1 File .

    Techniques Used:

    Drug induced effect on TER with respect to the rolling baseline. TER values at each stage of the drug regime chronologically are considered as baseline. The hypotenuse of the shaded triangular area represents the line of identity while magnification is shown in the box inserts. (A-I) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of forskolin (n = 44). (B-I) TER values +FSK are plotted against values after addition of CFTR Inh172 (n = 44). (C-I) Values of TER with CFTR Inh172 plotted against values +AMI (n = 44); line of regression: dashed line +99% CI (grey vertical bars). In D-I same data as in C-I showing two distinct populations of amiloride responders designated as LOW (closed grey circles-black dashed regression line +99% CI vertical bars, n = 35) and HIGH (open squares, black regression line +99% CI dotted line, n = 9). (A-II) The initial baseline TER values are plotted against the values obtained after addition of amiloride showing two distinct populations designated as LOW (n = 57) and HIGH (n = 24) amiloride responders. (B-II) Amiloride values plotted against values +FSK (LOW n = 56, HIGH n = 24). (C-II) Values after addition of forskolin plotted against values +CFTR Inh172 (LOW n = 56, HIGH n = 21). Additional regression and statistical analysis data are shown in Table C in S1 File .
    Figure Legend Snippet: Drug induced effect on TER with respect to the rolling baseline. TER values at each stage of the drug regime chronologically are considered as baseline. The hypotenuse of the shaded triangular area represents the line of identity while magnification is shown in the box inserts. (A-I) The initial baseline TER values (closed grey circles) are plotted against the values obtained after addition of forskolin (n = 44). (B-I) TER values +FSK are plotted against values after addition of CFTR Inh172 (n = 44). (C-I) Values of TER with CFTR Inh172 plotted against values +AMI (n = 44); line of regression: dashed line +99% CI (grey vertical bars). In D-I same data as in C-I showing two distinct populations of amiloride responders designated as LOW (closed grey circles-black dashed regression line +99% CI vertical bars, n = 35) and HIGH (open squares, black regression line +99% CI dotted line, n = 9). (A-II) The initial baseline TER values are plotted against the values obtained after addition of amiloride showing two distinct populations designated as LOW (n = 57) and HIGH (n = 24) amiloride responders. (B-II) Amiloride values plotted against values +FSK (LOW n = 56, HIGH n = 24). (C-II) Values after addition of forskolin plotted against values +CFTR Inh172 (LOW n = 56, HIGH n = 21). Additional regression and statistical analysis data are shown in Table C in S1 File .

    Techniques Used:

    37) Product Images from "Nedd4 mediates control of an epithelial Na+ channel in salivary duct cells by cytosolic Na+"

    Article Title: Nedd4 mediates control of an epithelial Na+ channel in salivary duct cells by cytosolic Na+

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    ( A ) The effects of the inclusion in NMDG-glutamate pipette solution of the GST–WW fusion protein (G-W), GST control (G), anti-Nedd4 IgG (A-Nd4), or pre-immune IgG on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( B ) The effects of the inclusion in 72 mmol/l Na + pipette solution of the GST control, the GST–WW fusion protein, pre-immune IgG, or anti-Nedd4 IgG on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( C ) The effects of the inclusion in NMDG–NO 3 pipette solution of the GST–WW fusion protein or anti-Nedd4 antibody on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( D ) The effects of the inclusion of activated α subunit of G o(αo) , activated α subunit of G o plus pre-immune IgG, activated α subunit of G o plus anti-Nedd4 IgG, or activated α subunit of G i2 plus anti-Nedd4 IgG in the NMDG-glutamate pipette solution on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. The chord conductance observed by using the NMDG-glutamate pipette solution is shown in each panel as a dotted line.
    Figure Legend Snippet: ( A ) The effects of the inclusion in NMDG-glutamate pipette solution of the GST–WW fusion protein (G-W), GST control (G), anti-Nedd4 IgG (A-Nd4), or pre-immune IgG on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( B ) The effects of the inclusion in 72 mmol/l Na + pipette solution of the GST control, the GST–WW fusion protein, pre-immune IgG, or anti-Nedd4 IgG on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( C ) The effects of the inclusion in NMDG–NO 3 pipette solution of the GST–WW fusion protein or anti-Nedd4 antibody on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. ( D ) The effects of the inclusion of activated α subunit of G o(αo) , activated α subunit of G o plus pre-immune IgG, activated α subunit of G o plus anti-Nedd4 IgG, or activated α subunit of G i2 plus anti-Nedd4 IgG in the NMDG-glutamate pipette solution on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance. The chord conductance observed by using the NMDG-glutamate pipette solution is shown in each panel as a dotted line.

    Techniques Used: Transferring

    The effects of the inclusion in the 72 mmol/l Na + pipette solution or in the NMDG-glutamate pipette solution of the GST–dn–ubiquitin (K48R) fusion protein (dn) and the GST–wt–ubiquitin fusion protein (wt) on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance.
    Figure Legend Snippet: The effects of the inclusion in the 72 mmol/l Na + pipette solution or in the NMDG-glutamate pipette solution of the GST–dn–ubiquitin (K48R) fusion protein (dn) and the GST–wt–ubiquitin fusion protein (wt) on the chord conductance measured at −80 mV of the amiloride-sensitive Na + conductance.

    Techniques Used: Transferring

    38) Product Images from "Feedback inhibition of rat amiloride-sensitive epithelial sodium channels expressed in Xenopus laevis oocytes"

    Article Title: Feedback inhibition of rat amiloride-sensitive epithelial sodium channels expressed in Xenopus laevis oocytes

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.1999.031aa.x

    Inhibition of ENaC downregulation during intracellular perfusion Original current recordings under cut-open oocyte conditions showing the decrease in I Amil when the membrane was clamped at -100 mV (downward deflections are due to voltage pulses to -60 mV). Filled bars above the current traces represent application of 5 μM amiloride. In A , the oocyte was impaled by the pipette but not perfused, the pipette being used only to record the intracellular voltage. The extracellular solution contained 50 mM Na + . B , when the intracellular side of the oocyte was perfused with a solution containing 20 mM Na + (extracellular [Na + ], 20 mM), I Amil remained stable over a 30-40 min period. C , a similar abolition of I Amil run-down was also seen when, after an initial 5-10 min perfusion with a 1 mM Na + intracellular solution (up to the time indicated by the arrow), the oocyte was perfused with a 50 mM Na + solution (extracellular [Na + ], 50 mM). No significant effect on I Amil or G Amil was observed following this increase in the Na + concentration of the intracellular perfusate.
    Figure Legend Snippet: Inhibition of ENaC downregulation during intracellular perfusion Original current recordings under cut-open oocyte conditions showing the decrease in I Amil when the membrane was clamped at -100 mV (downward deflections are due to voltage pulses to -60 mV). Filled bars above the current traces represent application of 5 μM amiloride. In A , the oocyte was impaled by the pipette but not perfused, the pipette being used only to record the intracellular voltage. The extracellular solution contained 50 mM Na + . B , when the intracellular side of the oocyte was perfused with a solution containing 20 mM Na + (extracellular [Na + ], 20 mM), I Amil remained stable over a 30-40 min period. C , a similar abolition of I Amil run-down was also seen when, after an initial 5-10 min perfusion with a 1 mM Na + intracellular solution (up to the time indicated by the arrow), the oocyte was perfused with a 50 mM Na + solution (extracellular [Na + ], 50 mM). No significant effect on I Amil or G Amil was observed following this increase in the Na + concentration of the intracellular perfusate.

    Techniques Used: Inhibition, Transferring, Concentration Assay

    Effect of intracellular perfusion: acidification A , effect on I Amil resulting from acidification of the intracellular perfusion solution from pH 7.4 to 6.5 (filled bars above the current trace represent application of 5 μM amiloride). At a flow rate of 5 μl min −1 , acidification decreased I Amil by about 40 %; I Amil reached a new steady state after about 2 min. When the pH was returned to control, in this example, I Amil reached about 85 % of the initial control current. The holding potential was -100 mV and the downward current deflections are due to the voltage steps to -60 mV used to monitor the membrane conductance. B , current-voltage relationships for I Amil before (•), during (○) and after (▪) a 3 min exposure to a pH 6.5 intracellular solution. In this example, the pH effect was fully reversible. The extracellular [Na + ] was 50 mM and the perfused [Na + ] was 1 mM.
    Figure Legend Snippet: Effect of intracellular perfusion: acidification A , effect on I Amil resulting from acidification of the intracellular perfusion solution from pH 7.4 to 6.5 (filled bars above the current trace represent application of 5 μM amiloride). At a flow rate of 5 μl min −1 , acidification decreased I Amil by about 40 %; I Amil reached a new steady state after about 2 min. When the pH was returned to control, in this example, I Amil reached about 85 % of the initial control current. The holding potential was -100 mV and the downward current deflections are due to the voltage steps to -60 mV used to monitor the membrane conductance. B , current-voltage relationships for I Amil before (•), during (○) and after (▪) a 3 min exposure to a pH 6.5 intracellular solution. In this example, the pH effect was fully reversible. The extracellular [Na + ] was 50 mM and the perfused [Na + ] was 1 mM.

    Techniques Used: Flow Cytometry

    Relationship between the apparent intracellular sodium concentration, [Na + ] i , and the amiloride-sensitive conductance ( G Amil ) of the exposed membrane The apparent [Na + ] i values were calculated from the reversal potential of the amiloride-sensitive current using a nominal external Na + concentration of 50 mM. All these values ( n = 10) were measured after a 30 min period during which the membrane potential was maintained at -100 mV and the intracellular side was continuously perfused with a 50 mM Na + solution. During this period, G Amil was stable (i.e. no run-down). The straight line is the linear regression for [Na + ] i versusG Amil and demonstrates a statistically significant correlation ( r 2 = 0.81, P
    Figure Legend Snippet: Relationship between the apparent intracellular sodium concentration, [Na + ] i , and the amiloride-sensitive conductance ( G Amil ) of the exposed membrane The apparent [Na + ] i values were calculated from the reversal potential of the amiloride-sensitive current using a nominal external Na + concentration of 50 mM. All these values ( n = 10) were measured after a 30 min period during which the membrane potential was maintained at -100 mV and the intracellular side was continuously perfused with a 50 mM Na + solution. During this period, G Amil was stable (i.e. no run-down). The straight line is the linear regression for [Na + ] i versusG Amil and demonstrates a statistically significant correlation ( r 2 = 0.81, P

    Techniques Used: Concentration Assay

    Current-voltage curves obtained using the cut-open oocyte technique A , current recordings obtained from a cut-open oocyte perfused with intracellular and extracellular solutions containing 50 mM Na + during a series of 175 ms square voltage pulses ranging from -140 to +60 mV. The exposed membrane (at the vegetal pole of the oocyte) had a diameter of ≈500 μm. B , current recordings as in A obtained after application of 5 μM amiloride. C , amiloride-sensitive currents (i.e. A - B ). D , current-voltage relationships for the whole-membrane current (•), residual current after application of amiloride (▪) and amiloride-sensitive current ( I Amil , ○). The currents were measured 150 ms after the beginning of the voltage pulse. V m , membrane potential.
    Figure Legend Snippet: Current-voltage curves obtained using the cut-open oocyte technique A , current recordings obtained from a cut-open oocyte perfused with intracellular and extracellular solutions containing 50 mM Na + during a series of 175 ms square voltage pulses ranging from -140 to +60 mV. The exposed membrane (at the vegetal pole of the oocyte) had a diameter of ≈500 μm. B , current recordings as in A obtained after application of 5 μM amiloride. C , amiloride-sensitive currents (i.e. A - B ). D , current-voltage relationships for the whole-membrane current (•), residual current after application of amiloride (▪) and amiloride-sensitive current ( I Amil , ○). The currents were measured 150 ms after the beginning of the voltage pulse. V m , membrane potential.

    Techniques Used: Mass Spectrometry

    39) Product Images from "Combined activity of COX-1 and COX-2 is increased in non-neoplastic colonic mucosa from colorectal neoplasia patients"

    Article Title: Combined activity of COX-1 and COX-2 is increased in non-neoplastic colonic mucosa from colorectal neoplasia patients

    Journal: BMC Gastroenterology

    doi: 10.1186/s12876-018-0759-1

    Examples of typical recording in Ussing Chamber experiments on short circuit current (SCC) following exposure to COX-1 (cyclooxygenase) inhibitor SC-560 ( a ) and COX-2 inhibitor celecoxib ( b ). Compounds were added to biopsies in the following concentrations: amiloride (20 μM, mucosal side), theophylline (400 μM, both sides), either COX-1 inhibitor (SC-560, 500 nM, both sides) or COX-2 inhibitor (celecoxib, 500 nM, both sides), indomethacin (13 μM, both sides) and prostaglandin (PGE 2 , 100 nM, serosal side)
    Figure Legend Snippet: Examples of typical recording in Ussing Chamber experiments on short circuit current (SCC) following exposure to COX-1 (cyclooxygenase) inhibitor SC-560 ( a ) and COX-2 inhibitor celecoxib ( b ). Compounds were added to biopsies in the following concentrations: amiloride (20 μM, mucosal side), theophylline (400 μM, both sides), either COX-1 inhibitor (SC-560, 500 nM, both sides) or COX-2 inhibitor (celecoxib, 500 nM, both sides), indomethacin (13 μM, both sides) and prostaglandin (PGE 2 , 100 nM, serosal side)

    Techniques Used:

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    other:

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes
    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Article Title: The K+ Channel Opener 1-EBIO Potentiates Residual Function of Mutant CFTR in Rectal Biopsies from Cystic Fibrosis Patients
    Article Snippet: Chemicals and Compounds Amiloride, indomethacin, carbachol, IBMX, forskolin, bumetanide and clotrimazole were all obtained from Sigma (Deisenhofen, Germany).

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    Millipore amiloride
    Effect of genistein on ion transport in rectal tissues from non-CF and CF individuals. (A) Concentration response curve for the effect of genistein (in the presence of 10 μmol l −1 indomethacin; basolateral side) on non-CF rectal mucosa (EC 50 =35 μmol l −1 ). (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF rectal mucosa. Experiments were performed in the presence of <t>amiloride</t> (10 μmol l −1 ; mucosal side) and in either(i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). # Indicates statistical significance for the effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect of genistein in the absence of cyclic AMP activation, in the presence of forskolin and in the presence of IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).
    Amiloride, 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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    Millipore human amidated npy
    Characterization of fluorogenic human amidated <t>NPY</t> . (A) Aliquots of unlabeled and <t>Alexa</t> Fluor‐labeled human amidated NPY were subjected to analysis by SDS / PAGE . To detect the peptide bands, gel was subjected to Coomassie Blue staining (Coomassie). The fluorescence of the labeled peptide was observed by placing the gel under UV light (Fluorescence). M, standard peptide marker. The positions of lower and upper bands of the labeled NPY are indicated by closed and opened arrowheads, respectively. (B) The labeled human amidated NPY was incubated with or without human native TTR at 37 °C, for 1 h. Then the reaction mixture was analyzed by SDS / PAGE .
    Human Amidated Npy, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore amiloride hydrochloride
    Use of the ATP scavenger hexokinase demonstrates the presence of separate apical and basolateral ATP receptors In these experiments hexokinase (126 units ml −1 ) was added to the apical ( A ) or basolateral ( B ) bath solution prior to sequential application of apical and basolateral ATP (100 μM). Apical <t>amiloride</t> (100 μM) was present throughout the experiment.
    Amiloride Hydrochloride, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore pglu
    Reaction scheme of DMTMM promoted amide coupling of <t>GlcN</t> and <t>PGlu</t> [ 64 , 68 ].
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    Effect of genistein on ion transport in rectal tissues from non-CF and CF individuals. (A) Concentration response curve for the effect of genistein (in the presence of 10 μmol l −1 indomethacin; basolateral side) on non-CF rectal mucosa (EC 50 =35 μmol l −1 ). (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF rectal mucosa. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either(i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). # Indicates statistical significance for the effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect of genistein in the absence of cyclic AMP activation, in the presence of forskolin and in the presence of IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Journal: British Journal of Pharmacology

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    doi: 10.1038/sj.bjp.0703520

    Figure Lengend Snippet: Effect of genistein on ion transport in rectal tissues from non-CF and CF individuals. (A) Concentration response curve for the effect of genistein (in the presence of 10 μmol l −1 indomethacin; basolateral side) on non-CF rectal mucosa (EC 50 =35 μmol l −1 ). (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF rectal mucosa. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either(i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). # Indicates statistical significance for the effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect of genistein in the absence of cyclic AMP activation, in the presence of forskolin and in the presence of IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Techniques: Concentration Assay, Activation Assay

    Effects of genistein on cholinergic secretion after deactivating (indomethacin) or activating (forskolin/IBMX) CFTR. Original recordings of V te obtained from rectal biopsies from (A) non-CF and (B) CF subjects. All experiments were performed in the presence of indomethacin (10 μmol l −1 ; basolateral side) and amiloride (10 μmol l −1 ; mucosal side). Cholinergic stimulation was induced by carbachol (CCH, 100 μmol l −1 ; basolateral side). (C) Summary of the CCH induced I sc in CF and non-CF rectal biopsies in the absence and presence of genistein (50 μmol l −1 mucosal side) and dependence on stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). #Indicates significant difference for CCH induced I sc in the presence of IBMX/forskolin compared to indomethacin (paired t -test). * Indicates statistical significance for the effect of genistein on CCH induced I sc in the presence of indomethacin and IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of genistein on CCH induced I sc in experiments obtained from non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Journal: British Journal of Pharmacology

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    doi: 10.1038/sj.bjp.0703520

    Figure Lengend Snippet: Effects of genistein on cholinergic secretion after deactivating (indomethacin) or activating (forskolin/IBMX) CFTR. Original recordings of V te obtained from rectal biopsies from (A) non-CF and (B) CF subjects. All experiments were performed in the presence of indomethacin (10 μmol l −1 ; basolateral side) and amiloride (10 μmol l −1 ; mucosal side). Cholinergic stimulation was induced by carbachol (CCH, 100 μmol l −1 ; basolateral side). (C) Summary of the CCH induced I sc in CF and non-CF rectal biopsies in the absence and presence of genistein (50 μmol l −1 mucosal side) and dependence on stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). #Indicates significant difference for CCH induced I sc in the presence of IBMX/forskolin compared to indomethacin (paired t -test). * Indicates statistical significance for the effect of genistein on CCH induced I sc in the presence of indomethacin and IBMX/forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of genistein on CCH induced I sc in experiments obtained from non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Techniques:

    Effect of genistein on ion transport in nasal epithelia from non-CF and CF subjects. (A) Concentration response curve for the effect of genistein (under resting conditions) in non-CF respiratory epithelia, showing an EC 50 of about 3.8 μmol l −1 . An inhibitory effect was observed at concentrations higher than 50 μmol l −1 . (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF nasal epithelia. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either (i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides). #Significant effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect genistein compared to control and in the presence of forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Journal: British Journal of Pharmacology

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    doi: 10.1038/sj.bjp.0703520

    Figure Lengend Snippet: Effect of genistein on ion transport in nasal epithelia from non-CF and CF subjects. (A) Concentration response curve for the effect of genistein (under resting conditions) in non-CF respiratory epithelia, showing an EC 50 of about 3.8 μmol l −1 . An inhibitory effect was observed at concentrations higher than 50 μmol l −1 . (B) Summary of the effects of 50 μmol l −1 genistein (mucosal side) on non-CF and CF nasal epithelia. Experiments were performed in the presence of amiloride (10 μmol l −1 ; mucosal side) and in either (i) absence of cyclic AMP dependent stimulation, (ii) after submaximal stimulation with low concentrations of forskolin (0.1 μmol l −1 ; basolateral side) or (iii) after maximal stimulation with IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides). #Significant effects of forskolin and IBMX/forskolin compared to control (paired t -test). * Indicates statistical significance for the effect genistein compared to control and in the presence of forskolin (paired t -test). ≈rcub;Indicates statistical significance when comparing the effect of genistein under the different experimental conditions (see above) in non-CF and CF tissues (unpaired t -test). (Number of subjects).

    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Techniques: Concentration Assay

    (A) Summary of short circuit currents (I sc ) in rectal epithelia from non-CF and CF individuals under resting conditions, after incubation with indomethacin (10 μmol l −1 ; basolateral side), after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). * Indicates statistical significance for the effects of indomethacin, amiloride and IBMX/forskolin in non-CF or CF (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of indomethacin, amiloride and IBMX/forskolin obtained in experiments from rectal tissues of non-CF and CF subjects (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human rectal mucosa. The effects of genistein were examined in both absence and presence of IBMX and forskolin and experiments were carried out in the presence of indomethacin and amiloride. R te was continuously determined from the V te downward deflections obtained by pulsed current injection.

    Journal: British Journal of Pharmacology

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    doi: 10.1038/sj.bjp.0703520

    Figure Lengend Snippet: (A) Summary of short circuit currents (I sc ) in rectal epithelia from non-CF and CF individuals under resting conditions, after incubation with indomethacin (10 μmol l −1 ; basolateral side), after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; basolateral side). * Indicates statistical significance for the effects of indomethacin, amiloride and IBMX/forskolin in non-CF or CF (paired t -test). ≈rcub;Indicates statistical significance when comparing the effects of indomethacin, amiloride and IBMX/forskolin obtained in experiments from rectal tissues of non-CF and CF subjects (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human rectal mucosa. The effects of genistein were examined in both absence and presence of IBMX and forskolin and experiments were carried out in the presence of indomethacin and amiloride. R te was continuously determined from the V te downward deflections obtained by pulsed current injection.

    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Techniques: Incubation, Blocking Assay, Activation Assay, Injection

    (A) Summary of short circuit currents (I sc ) under resting conditions, after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides) in human nasal epithelia from non-CF and CF individuals. * Indicates statistical significance for the effects of amiloride and IBMX/forskolin (paired t -test). §Indicate statistical significance for the basal I sc and the effects of amiloride and IBMX/forskolin when compared to experiments obtained from non-CF tissues (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human nasal tissues. Epithelial Na + conductance was blocked by amiloride and effects of genistein were examined in both absence and presence of IBMX and forskolin. R te was determined continuously from the V te downward deflections obtained by pulsed current injection.

    Journal: British Journal of Pharmacology

    Article Title: Effect of genistein on native epithelial tissue from normal individuals and CF patients and on ion channels expressed in Xenopus oocytes

    doi: 10.1038/sj.bjp.0703520

    Figure Lengend Snippet: (A) Summary of short circuit currents (I sc ) under resting conditions, after blocking epithelial Na + channels by amiloride (10 μmol l −1 ; mucosal side) and after activation of ion transport by IBMX and forskolin (100 μmol l −1 and 1 μmol l −1 ; both sides) in human nasal epithelia from non-CF and CF individuals. * Indicates statistical significance for the effects of amiloride and IBMX/forskolin (paired t -test). §Indicate statistical significance for the basal I sc and the effects of amiloride and IBMX/forskolin when compared to experiments obtained from non-CF tissues (unpaired t -test). (Number of subjects). Effect of genistein (50 μmol l −1 ; mucosal side) on transepithelial voltage (V te ) in (B) non-CF and (C) CF human nasal tissues. Epithelial Na + conductance was blocked by amiloride and effects of genistein were examined in both absence and presence of IBMX and forskolin. R te was determined continuously from the V te downward deflections obtained by pulsed current injection.

    Article Snippet: Amiloride, indomethacin, genistein and IBMX were all obtained from Sigma (Deisenhofen, Germany).

    Techniques: Blocking Assay, Activation Assay, Injection

    Characterization of fluorogenic human amidated NPY . (A) Aliquots of unlabeled and Alexa Fluor‐labeled human amidated NPY were subjected to analysis by SDS / PAGE . To detect the peptide bands, gel was subjected to Coomassie Blue staining (Coomassie). The fluorescence of the labeled peptide was observed by placing the gel under UV light (Fluorescence). M, standard peptide marker. The positions of lower and upper bands of the labeled NPY are indicated by closed and opened arrowheads, respectively. (B) The labeled human amidated NPY was incubated with or without human native TTR at 37 °C, for 1 h. Then the reaction mixture was analyzed by SDS / PAGE .

    Journal: FEBS Open Bio

    Article Title: The hydrophobic C‐terminal sequence of transthyretin affects its catalytic kinetics towards amidated neuropeptide Y

    doi: 10.1002/2211-5463.12604

    Figure Lengend Snippet: Characterization of fluorogenic human amidated NPY . (A) Aliquots of unlabeled and Alexa Fluor‐labeled human amidated NPY were subjected to analysis by SDS / PAGE . To detect the peptide bands, gel was subjected to Coomassie Blue staining (Coomassie). The fluorescence of the labeled peptide was observed by placing the gel under UV light (Fluorescence). M, standard peptide marker. The positions of lower and upper bands of the labeled NPY are indicated by closed and opened arrowheads, respectively. (B) The labeled human amidated NPY was incubated with or without human native TTR at 37 °C, for 1 h. Then the reaction mixture was analyzed by SDS / PAGE .

    Article Snippet: To label human amidated NPY with Alexa Fluor 488, we followed the standard protocol suggested by the manufacturer at pH 8.0.

    Techniques: Labeling, SDS Page, Staining, Fluorescence, Marker, Incubation

    Use of the ATP scavenger hexokinase demonstrates the presence of separate apical and basolateral ATP receptors In these experiments hexokinase (126 units ml −1 ) was added to the apical ( A ) or basolateral ( B ) bath solution prior to sequential application of apical and basolateral ATP (100 μM). Apical amiloride (100 μM) was present throughout the experiment.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Use of the ATP scavenger hexokinase demonstrates the presence of separate apical and basolateral ATP receptors In these experiments hexokinase (126 units ml −1 ) was added to the apical ( A ) or basolateral ( B ) bath solution prior to sequential application of apical and basolateral ATP (100 μM). Apical amiloride (100 μM) was present throughout the experiment.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Pretreatment with extracellular BAPTA AM blunted the response to apical ATP A, control response to apical ATP (100 μM). B, response to apical ATP (100 μM) in cells pretreated with 50 μM BAPTA AM for 60 min. Amiloride (100 μM) was present throughout the experiment and 1 mM DPC was added apically as indicated.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Pretreatment with extracellular BAPTA AM blunted the response to apical ATP A, control response to apical ATP (100 μM). B, response to apical ATP (100 μM) in cells pretreated with 50 μM BAPTA AM for 60 min. Amiloride (100 μM) was present throughout the experiment and 1 mM DPC was added apically as indicated.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Effect of apical ATP on the I SC of M-1 mouse CCD cells A, continuous I SC recording in a control experiment demonstrating long term stability of I SC and the effect of apical (ap) amiloride (100 μM) and DPC (1 mM) on resting I SC . Drugs were applied as indicated by the horizontal bars. Positive I SC corresponds to electrogenic cation absorption or anion secretion or a combination of both. B, effect of apical application of ATP (100 μM) on I SC of M-1 cells from the same batch of cells as in A ; amiloride (100 μM) and DPC (1 mM) were apically applied in the continuous presence of ATP.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Effect of apical ATP on the I SC of M-1 mouse CCD cells A, continuous I SC recording in a control experiment demonstrating long term stability of I SC and the effect of apical (ap) amiloride (100 μM) and DPC (1 mM) on resting I SC . Drugs were applied as indicated by the horizontal bars. Positive I SC corresponds to electrogenic cation absorption or anion secretion or a combination of both. B, effect of apical application of ATP (100 μM) on I SC of M-1 cells from the same batch of cells as in A ; amiloride (100 μM) and DPC (1 mM) were apically applied in the continuous presence of ATP.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Apical Cl − channel inhibitors significantly reduced the apical I SC response to ATP Online recording of the I SC responses to apical ATP (100 μM) before, during and after apical application of 1 mM DPC ( A ), 300 μM DIDS ( B ) and 100 μM niflumic acid ( C ). Apical amiloride (100 μM) was present throughout.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Apical Cl − channel inhibitors significantly reduced the apical I SC response to ATP Online recording of the I SC responses to apical ATP (100 μM) before, during and after apical application of 1 mM DPC ( A ), 300 μM DIDS ( B ) and 100 μM niflumic acid ( C ). Apical amiloride (100 μM) was present throughout.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Removal of extracellular Cl − inhibited the response to apical ATP M-1 cells were exposed to apical ATP (100 μM) in the presence and absence of extracellular Cl − . The experiment was conducted in the continuous presence of 100 μM amiloride.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Removal of extracellular Cl − inhibited the response to apical ATP M-1 cells were exposed to apical ATP (100 μM) in the presence and absence of extracellular Cl − . The experiment was conducted in the continuous presence of 100 μM amiloride.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Basolateral ATP has a similar effect to apical ATP Corresponding traces of I SC ( A ), V te ( B ) and R te ( C ) from an experiment during which ATP (100 μM) was added basolaterally (bl) in the absence and presence of apical amiloride (100 μM) are shown. The noisy I SC trace after the first ATP application is due to washout of ATP.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: Basolateral ATP has a similar effect to apical ATP Corresponding traces of I SC ( A ), V te ( B ) and R te ( C ) from an experiment during which ATP (100 μM) was added basolaterally (bl) in the absence and presence of apical amiloride (100 μM) are shown. The noisy I SC trace after the first ATP application is due to washout of ATP.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    The I SC response to ATP is preserved in the presence of amiloride M-1 cells were exposed to apical ATP (100 μM) in the absence and presence of apical amiloride (100 μM) as indicated by the horizontal bars. The noisy I SC trace after the first ATP application is due to washout of ATP.

    Journal: The Journal of Physiology

    Article Title: ATP stimulates Cl− secretion and reduces amiloride-sensitive Na+ absorption in M-1 mouse cortical collecting duct cells

    doi: 10.1111/j.1469-7793.2000.00077.x

    Figure Lengend Snippet: The I SC response to ATP is preserved in the presence of amiloride M-1 cells were exposed to apical ATP (100 μM) in the absence and presence of apical amiloride (100 μM) as indicated by the horizontal bars. The noisy I SC trace after the first ATP application is due to washout of ATP.

    Article Snippet: Amiloride hydrochloride, adenosine 5′-triphosphate (ATP) disodium salt, adenosine 5′-diphosphate (ADP) sodium salt, adenosine 5′-monophosphate (AMP) sodium salt, uridine 5′-triphosphate (UTP) sodium salt, uridine 5′-diphosphate (UDP) sodium salt, 2-methylthioATP (2-MeSATP) tetrasodium salt, bumetanide, 4,4′-diisothiocyanato-stilbene-2,2′-disulphonic acid disodium salt (DIDS), niflumic acid and hexokinase were purchased from Sigma-Aldrich (Steinheim, Germany), diphenylamine-2-carboxylic acid (DPC) was obtained from Fluka (Neu-Ulm, Germany).

    Techniques:

    Reaction scheme of DMTMM promoted amide coupling of GlcN and PGlu [ 64 , 68 ].

    Journal: Molecules

    Article Title: Post-Polymerization Modification of Poly(l-glutamic acid) with d-(+)-Glucosamine

    doi: 10.3390/molecules191219751

    Figure Lengend Snippet: Reaction scheme of DMTMM promoted amide coupling of GlcN and PGlu [ 64 , 68 ].

    Article Snippet: Two different eluents were used: (i) 0.1 M sodium nitrate (NaNO3 ) solution with sodium azide (NaN3 ) (0.02% w/v) (both from Sigma-Aldrich), prepared with MiliQ water (18.2 MΩ/cm) at pH 10 for PGlu and P(Glu-GlcN) samples; and (ii) N ,N -dimethylacetamide (DMAc) with 0.05 M lithium bromide (LiBr) (both from Sigma-Aldrich) for PBGlu sample.

    Techniques: