Structured Review

Millipore dids
Effects of anion channel blockers on I Cl,pH . (A, B) Representative current traces obtained by step-like pulses, same as in . I Cl,pH was activated by pH e 5.0, and <t>DIDS</t> (1 µM) or <t>niflumic</t> acid (NFA, 100 µM) was added (right panels).
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Images

1) Product Images from "Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes"

Article Title: Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes

Journal:

doi: 10.4196/kjpp.2008.12.4.177

Effects of anion channel blockers on I Cl,pH . (A, B) Representative current traces obtained by step-like pulses, same as in . I Cl,pH was activated by pH e 5.0, and DIDS (1 µM) or niflumic acid (NFA, 100 µM) was added (right panels).
Figure Legend Snippet: Effects of anion channel blockers on I Cl,pH . (A, B) Representative current traces obtained by step-like pulses, same as in . I Cl,pH was activated by pH e 5.0, and DIDS (1 µM) or niflumic acid (NFA, 100 µM) was added (right panels).

Techniques Used:

2) Product Images from "Molecular determinants of differential pore blocking of kidney CLC-K chloride channels"

Article Title: Molecular determinants of differential pore blocking of kidney CLC-K chloride channels

Journal: EMBO Reports

doi: 10.1038/sj.embor.7400169

Effects of 3-phenyl-CPP and DIDS on CLC-Kb. Voltage-clamp traces of CLC-Kb currents before and during application of 200 μM 3-phenyl-CPP ( A ) and 200 μM DIDS ( B ), and immediately after wash-out. Horizontal scale bars indicate 100 ms. Vertical scale bars indicate 2 μA ( A ) and 1 μA ( B ).
Figure Legend Snippet: Effects of 3-phenyl-CPP and DIDS on CLC-Kb. Voltage-clamp traces of CLC-Kb currents before and during application of 200 μM 3-phenyl-CPP ( A ) and 200 μM DIDS ( B ), and immediately after wash-out. Horizontal scale bars indicate 100 ms. Vertical scale bars indicate 2 μA ( A ) and 1 μA ( B ).

Techniques Used: Conditioned Place Preference, Mass Spectrometry

Effects of 3-phenyl-CPP and DIDS on CLC-Ka. ( A,B ) Voltage-clamp traces of CLC-Ka currents before and during application of 200 μM 3-phenyl-CPP ( A ) and 200 μM DIDS ( B ), and immediately after wash-out. The pulse protocol is shown in the inset. Horizontal scale bars indicate 50 ms ( A,B ). Vertical scale bars indicate 5 μA ( A,B ). ( C ) Chemical structures of 3-phenyl-CPP and DIDS. ( D ) Slow recovery from DIDS block for a representative oocyte. Arrows indicate application of DIDS and wash-out. A pulse to 60 mV was applied every 2 s and the current is shown as a function of the elapsed time.
Figure Legend Snippet: Effects of 3-phenyl-CPP and DIDS on CLC-Ka. ( A,B ) Voltage-clamp traces of CLC-Ka currents before and during application of 200 μM 3-phenyl-CPP ( A ) and 200 μM DIDS ( B ), and immediately after wash-out. The pulse protocol is shown in the inset. Horizontal scale bars indicate 50 ms ( A,B ). Vertical scale bars indicate 5 μA ( A,B ). ( C ) Chemical structures of 3-phenyl-CPP and DIDS. ( D ) Slow recovery from DIDS block for a representative oocyte. Arrows indicate application of DIDS and wash-out. A pulse to 60 mV was applied every 2 s and the current is shown as a function of the elapsed time.

Techniques Used: Conditioned Place Preference, Mass Spectrometry, Blocking Assay

3) Product Images from "An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons"

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons

Journal: Brain Research

doi: 10.1016/j.brainres.2012.03.004

DIDS penetrates damaged, but not intact neuronal membranes (A) Two-photon confocal DIC and Z-stack projection images (2 microns total) of neurons treated without DIDS, or with DIDS during normoxia or IS. Upper panel: DIC images. Lower panel: DIC images (grey) overlaid with PI (red) and DIDS (blue) fluorescent Z-stack projection images. Images are representative of 4 separate experiments for each treatment. (B) Summary of PI uptake. Data are mean ± SEM from 10–20 replicates per treatment. Asterisks (*) indicate significant difference from control ( p
Figure Legend Snippet: DIDS penetrates damaged, but not intact neuronal membranes (A) Two-photon confocal DIC and Z-stack projection images (2 microns total) of neurons treated without DIDS, or with DIDS during normoxia or IS. Upper panel: DIC images. Lower panel: DIC images (grey) overlaid with PI (red) and DIDS (blue) fluorescent Z-stack projection images. Images are representative of 4 separate experiments for each treatment. (B) Summary of PI uptake. Data are mean ± SEM from 10–20 replicates per treatment. Asterisks (*) indicate significant difference from control ( p

Techniques Used:

4) Product Images from "Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism"

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism

Journal: Scientific Reports

doi: 10.1038/srep31433

Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p
Figure Legend Snippet: Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p

Techniques Used:

5) Product Images from "Giardia secretome highlights secreted tenascins as a key component of pathogenesis"

Article Title: Giardia secretome highlights secreted tenascins as a key component of pathogenesis

Journal: GigaScience

doi: 10.1093/gigascience/giy003

The effect of co-culture with Giardia or Giardia supernatants on the electrophysiological properties of CaCo-2 monolayers. (A) Transepithelial electrical resistance in CaCo-2 monolayers following seeding on permeable supports. Data show an increase in TEER as the monolayer develops. Confluence occurred around day 6. Giardia were added on day 6 after the confluent monolayer formed and co-cultured with the Caco-2 monolayer for 24 hours. TEER was measured after 24 hours and compared with TEER in monolayers that had not been exposed to Giardia (n = 6). (B) A representative short circuit current (Isc) against time recording from single monolayers of CaCo-2 cells in an Ussing chamber. The trace shows the activation of CFTR chloride channels (basolateral application of 10 μM of forskolin) and calcium-activated chloride channels (basolateral application of 100 μM of UTP). Specificity of activation is confirmed by inhibition of Isc by the specific CFTR channel blocker, GlyH101; and specific calcium-activated chloride channel blocker, DIDS. The effect on Isc of 24-hour co-incubation of CaCo-2 monolayers with Giardia or with Giardia supernatant (1:1000 dilution) is also shown. (C) Effect of 24-hour co-incubation of CaCo-2 monolayers with different strains of Giardia (WB, GS, and patient samples) on forskolin-stimulated and UTP-stimulated Isc (n = 3). (D) Effect of supernatant co-incubation from different strains of Giardia (WB, GS, and patient samples) on forskolin-stimulated and UTP-stimulated Isc (n = 3) from Caco-2 monolayers. The results were analysed by the Student t test and expressed as mean values ± standard error mean (SEM). Significant difference expressed as * P
Figure Legend Snippet: The effect of co-culture with Giardia or Giardia supernatants on the electrophysiological properties of CaCo-2 monolayers. (A) Transepithelial electrical resistance in CaCo-2 monolayers following seeding on permeable supports. Data show an increase in TEER as the monolayer develops. Confluence occurred around day 6. Giardia were added on day 6 after the confluent monolayer formed and co-cultured with the Caco-2 monolayer for 24 hours. TEER was measured after 24 hours and compared with TEER in monolayers that had not been exposed to Giardia (n = 6). (B) A representative short circuit current (Isc) against time recording from single monolayers of CaCo-2 cells in an Ussing chamber. The trace shows the activation of CFTR chloride channels (basolateral application of 10 μM of forskolin) and calcium-activated chloride channels (basolateral application of 100 μM of UTP). Specificity of activation is confirmed by inhibition of Isc by the specific CFTR channel blocker, GlyH101; and specific calcium-activated chloride channel blocker, DIDS. The effect on Isc of 24-hour co-incubation of CaCo-2 monolayers with Giardia or with Giardia supernatant (1:1000 dilution) is also shown. (C) Effect of 24-hour co-incubation of CaCo-2 monolayers with different strains of Giardia (WB, GS, and patient samples) on forskolin-stimulated and UTP-stimulated Isc (n = 3). (D) Effect of supernatant co-incubation from different strains of Giardia (WB, GS, and patient samples) on forskolin-stimulated and UTP-stimulated Isc (n = 3) from Caco-2 monolayers. The results were analysed by the Student t test and expressed as mean values ± standard error mean (SEM). Significant difference expressed as * P

Techniques Used: Co-Culture Assay, Cell Culture, Activation Assay, Inhibition, Incubation, Western Blot

6) Product Images from "The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility"

Article Title: The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.RA117.001343

Cl − channel blockers inhibit the metastatic phenotypic changes associated with ST expression. A and B , in ( A ) i293-EGFP and i293-EGFP-ST cells were induced with doxycycline hyclate and in ( B ) MCC13 cells were transfected with EGFP and EGFP-ST. ( i ) cells were then treated with DMSO, DIDS, RIAA, or NPPB and after 24 h, cell motility was measured using the IncuCyte kinetic imaging system. Images were taken every 30 min for a 24-h period. ( ii ) scatter plot showing cell movement tracked using ImageJ ( n = 25). Average cell movement, as indicated by the horizontal bar , was calculated and significance tested using a two-tailed Student's t test; *, p
Figure Legend Snippet: Cl − channel blockers inhibit the metastatic phenotypic changes associated with ST expression. A and B , in ( A ) i293-EGFP and i293-EGFP-ST cells were induced with doxycycline hyclate and in ( B ) MCC13 cells were transfected with EGFP and EGFP-ST. ( i ) cells were then treated with DMSO, DIDS, RIAA, or NPPB and after 24 h, cell motility was measured using the IncuCyte kinetic imaging system. Images were taken every 30 min for a 24-h period. ( ii ) scatter plot showing cell movement tracked using ImageJ ( n = 25). Average cell movement, as indicated by the horizontal bar , was calculated and significance tested using a two-tailed Student's t test; *, p

Techniques Used: Expressing, Transfection, Imaging, Two Tailed Test

7) Product Images from "The Interaction between Trolox and 4,4’-diisothiocyanatostilbene-2,2’-disulfonic Acid on Hypoxic Pulmonary Vasoconstriction in the Isolated Rabbit Lung"

Article Title: The Interaction between Trolox and 4,4’-diisothiocyanatostilbene-2,2’-disulfonic Acid on Hypoxic Pulmonary Vasoconstriction in the Isolated Rabbit Lung

Journal: Iranian Journal of Medical Sciences

doi:

Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups were lower than other groups at 30 minutes. Data are presented as mean±SE (n=5 in each group). **P
Figure Legend Snippet: Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups were lower than other groups at 30 minutes. Data are presented as mean±SE (n=5 in each group). **P

Techniques Used:

Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups were lower than other groups at 5 minutes. Data are presented as mean±SE (n=5 in each group). *P
Figure Legend Snippet: Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups were lower than other groups at 5 minutes. Data are presented as mean±SE (n=5 in each group). *P

Techniques Used:

Concentrations of NO metabolites of the perfusate in Trolox+DIDS-treated was more than other groups at 30 minutes. Data are presented as mean±SE (n=5 in each group). **P
Figure Legend Snippet: Concentrations of NO metabolites of the perfusate in Trolox+DIDS-treated was more than other groups at 30 minutes. Data are presented as mean±SE (n=5 in each group). **P

Techniques Used:

Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups at 30 minutes were lower than their values at 5 minutes. Data are presented as mean±SE (n=5 in each group). *P
Figure Legend Snippet: Pulmonary vascular resistance (∆PVR) in Trolox-treated and Trolox+DIDS-treated groups at 30 minutes were lower than their values at 5 minutes. Data are presented as mean±SE (n=5 in each group). *P

Techniques Used:

8) Product Images from "The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility"

Article Title: The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.RA117.001343

Cl − channel blockers inhibit the metastatic phenotypic changes associated with ST expression. A and B , in ( A ) i293-EGFP and i293-EGFP-ST cells were induced with doxycycline hyclate and in ( B ) MCC13 cells were transfected with EGFP and EGFP-ST. ( i ) cells were then treated with DMSO, DIDS, RIAA, or NPPB and after 24 h, cell motility was measured using the IncuCyte kinetic imaging system. Images were taken every 30 min for a 24-h period. ( ii ) scatter plot showing cell movement tracked using ImageJ ( n = 25). Average cell movement, as indicated by the horizontal bar , was calculated and significance tested using a two-tailed Student's t test; *, p
Figure Legend Snippet: Cl − channel blockers inhibit the metastatic phenotypic changes associated with ST expression. A and B , in ( A ) i293-EGFP and i293-EGFP-ST cells were induced with doxycycline hyclate and in ( B ) MCC13 cells were transfected with EGFP and EGFP-ST. ( i ) cells were then treated with DMSO, DIDS, RIAA, or NPPB and after 24 h, cell motility was measured using the IncuCyte kinetic imaging system. Images were taken every 30 min for a 24-h period. ( ii ) scatter plot showing cell movement tracked using ImageJ ( n = 25). Average cell movement, as indicated by the horizontal bar , was calculated and significance tested using a two-tailed Student's t test; *, p

Techniques Used: Expressing, Transfection, Imaging, Two Tailed Test

9) Product Images from "PI3K and PKC contribute to membrane depolarization mediated by ?2-adrenoceptors in the canine isolated mesenteric vein"

Article Title: PI3K and PKC contribute to membrane depolarization mediated by ?2-adrenoceptors in the canine isolated mesenteric vein

Journal: BMC Physiology

doi: 10.1186/1472-6793-5-9

The EFS-induced slow membrane depolarization of canine isolated mesenteric vein is reduced by various ion channel inhibitors . Intracellular recordings of membrane potential in response to EFS (0.1 ms, 13 V, 10 s, 0.5 Hz) in canine isolated mesenteric veins preincubated for 60 min prior to and throughout the experiment with 100 μM NFA (A), 30 μM NPPB (B), 200 μM DIDS (C) and 30 μM Gd 3+ (D). The experimental data are presented as mean ± SEM, n = 3–4 (E). *, P
Figure Legend Snippet: The EFS-induced slow membrane depolarization of canine isolated mesenteric vein is reduced by various ion channel inhibitors . Intracellular recordings of membrane potential in response to EFS (0.1 ms, 13 V, 10 s, 0.5 Hz) in canine isolated mesenteric veins preincubated for 60 min prior to and throughout the experiment with 100 μM NFA (A), 30 μM NPPB (B), 200 μM DIDS (C) and 30 μM Gd 3+ (D). The experimental data are presented as mean ± SEM, n = 3–4 (E). *, P

Techniques Used: Isolation, Mass Spectrometry

10) Product Images from "Sat1 is dispensable for active oxalate secretion in mouse duodenum"

Article Title: Sat1 is dispensable for active oxalate secretion in mouse duodenum

Journal: American Journal of Physiology - Cell Physiology

doi: 10.1152/ajpcell.00385.2011

Basolateral DIDS inhibits oxalate secretion. Transepithelial secretory fluxes of [ 14 C]oxalate and [ 3 H]mannitol were measured simultaneously across wild-type (WT) mouse duodenum in the absence and presence of 1 mM DIDS added to the serosal solution. P app , apparent permeability coefficient. Values are means ± SE ( n = 4). * P value
Figure Legend Snippet: Basolateral DIDS inhibits oxalate secretion. Transepithelial secretory fluxes of [ 14 C]oxalate and [ 3 H]mannitol were measured simultaneously across wild-type (WT) mouse duodenum in the absence and presence of 1 mM DIDS added to the serosal solution. P app , apparent permeability coefficient. Values are means ± SE ( n = 4). * P value

Techniques Used: Permeability

11) Product Images from "SCRAMBLING OF PHOSPHOLIPIDS ACTIVATES RED CELL MEMBRANE CHOLESTEROL"

Article Title: SCRAMBLING OF PHOSPHOLIPIDS ACTIVATES RED CELL MEMBRANE CHOLESTEROL

Journal: Biochemistry

doi: 10.1021/bi6023397

Effect of scramblase inhibitors on ionomycin-Ca ++ promoted oxidation of red cell cholesterol. Washed cells (80 μl) were incubated for 30 min at 37 °C in 0.72 ml PBS, 25 μM DIDS or 1.5 mM TNBS, all containing 0.1 mM EGTA. The cells
Figure Legend Snippet: Effect of scramblase inhibitors on ionomycin-Ca ++ promoted oxidation of red cell cholesterol. Washed cells (80 μl) were incubated for 30 min at 37 °C in 0.72 ml PBS, 25 μM DIDS or 1.5 mM TNBS, all containing 0.1 mM EGTA. The cells

Techniques Used: Incubation

12) Product Images from "Exploring the role of stromal osmoregulation in cancer and disease using executable modelling"

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling

Journal: Nature Communications

doi: 10.1038/s41467-018-05414-y

Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P
Figure Legend Snippet: Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P

Techniques Used: In Vitro, Cell Attachment Assay

13) Product Images from "Chloride channel blocker 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid inhibits nitric oxide-induced apoptosis in cultured rat hippocampal neurons ★"

Article Title: Chloride channel blocker 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid inhibits nitric oxide-induced apoptosis in cultured rat hippocampal neurons ★

Journal: Neural Regeneration Research

doi: 10.3969/j.issn.1673-5374.2013.02.003

Immunofluorescence staining of apoptosis-inducing factor (AIF) in rat hippocampal neurons (inverted fluorescence microscope, × 200). The specific neuronal antibody anti-NeuN was green (FITC labeled), AIF expression was significantly increased in the SIN-1 group (Cy3 labeled), and significantly decreased after treatment with the chloride channel blocker DIDS. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.
Figure Legend Snippet: Immunofluorescence staining of apoptosis-inducing factor (AIF) in rat hippocampal neurons (inverted fluorescence microscope, × 200). The specific neuronal antibody anti-NeuN was green (FITC labeled), AIF expression was significantly increased in the SIN-1 group (Cy3 labeled), and significantly decreased after treatment with the chloride channel blocker DIDS. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.

Techniques Used: Immunofluorescence, Staining, Fluorescence, Microscopy, Labeling, Expressing

Apoptotic neurons in rat hippocampus. (A) Morphology of cultured rat hippocampal neurons following DNA fluorescent staining (Hoechst 33258 staining, inverted fluorescence microscope, × 200). Arrows indicate apoptotic neurons (Hoechst 33258- positive cells displayed strong blue fluorescence). Apoptotic cells were evident in the 3-morpholinosydnonimine (SIN-1; 1.0 mM) group, but fewer were visible in the SIN-1 + DIDS group. (B) Quantification of apoptotic cells. a P
Figure Legend Snippet: Apoptotic neurons in rat hippocampus. (A) Morphology of cultured rat hippocampal neurons following DNA fluorescent staining (Hoechst 33258 staining, inverted fluorescence microscope, × 200). Arrows indicate apoptotic neurons (Hoechst 33258- positive cells displayed strong blue fluorescence). Apoptotic cells were evident in the 3-morpholinosydnonimine (SIN-1; 1.0 mM) group, but fewer were visible in the SIN-1 + DIDS group. (B) Quantification of apoptotic cells. a P

Techniques Used: Cell Culture, Staining, Fluorescence, Microscopy

Expression of poly(adenosine diphosphate- ribose)polymerase-1 (PARP-1) in rat hippocampal neurons (inverted fluorescence microscope, × 200). The specific neuronal antibody anti-NeuN was green (FITC labeled) and PARP-1 was red (Cy3 labeled). PARP-1 expression was significantly increased in the SIN-1 group, and was significantly decreased after treatment with the chloride channel blocker DIDS. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.
Figure Legend Snippet: Expression of poly(adenosine diphosphate- ribose)polymerase-1 (PARP-1) in rat hippocampal neurons (inverted fluorescence microscope, × 200). The specific neuronal antibody anti-NeuN was green (FITC labeled) and PARP-1 was red (Cy3 labeled). PARP-1 expression was significantly increased in the SIN-1 group, and was significantly decreased after treatment with the chloride channel blocker DIDS. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.

Techniques Used: Expressing, Fluorescence, Microscopy, Labeling

Poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1) and apoptosis-inducing factor (AIF) protein expression in rat hippocampal neurons. Expression levels in the SIN-1 group were significantly higher than those in the control group. DIDS downregulated PARP-1 and AIF expression. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.
Figure Legend Snippet: Poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1) and apoptosis-inducing factor (AIF) protein expression in rat hippocampal neurons. Expression levels in the SIN-1 group were significantly higher than those in the control group. DIDS downregulated PARP-1 and AIF expression. SIN-1: 3-morpholinosydnonimine; DIDS: 4,4-diisothiocyanatostilbene-2,2′-disulfonic acid.

Techniques Used: Expressing

14) Product Images from "Downregulation of LRRC8A protects human ovarian and alveolar carcinoma cells against Cisplatin-induced expression of p53, MDM2, p21Waf1/Cip1, and Caspase-9/-3 activation"

Article Title: Downregulation of LRRC8A protects human ovarian and alveolar carcinoma cells against Cisplatin-induced expression of p53, MDM2, p21Waf1/Cip1, and Caspase-9/-3 activation

Journal: American Journal of Physiology - Cell Physiology

doi: 10.1152/ajpcell.00256.2015

Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures ). A : A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B : A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t -test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D : A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).
Figure Legend Snippet: Cotreatment with anion-channel blockers and transient knock-down of LRRC8A abolish Cisplatin-induced but not TNFα and Hyperosmotic-induced Caspase-3 activation in wild-type A2780 cells. A2780WT and A2780CisR cell lysates were used for Caspase-3 activity assay using a commercial kit (see experimental procedures ). A : A2780CisR cells were exposed to 10 μM Cisplatin for 18 h and A2780WT cells were likewise treated with Cisplatin in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (A2780WT) and 4 (A2780CisR) individual sets of experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in the absence of the inhibitor, respectively (ANOVA, Fisher LSD method). B : A2780WT were treated with either 25 nM scramble siRNA or LRRC8A siRNA for 30 h and subsequently exposed to 10 μM Cisplatin for another 18 h. Data represent 4 individual experiments ± SE. * and #: Significantly increased by Cisplatin compared with the respective control and significantly reduced compared with Cisplatin treatment in Scramble siRNA-treated cells (Student's t -test). C: A2780CisR cells were exposed to 20 nM TNFα for 18 h. A2780WT cells were likewise treated with TNFα in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 6–8 (WT) and 4 (CisR) individual experiments ± SE. *Significantly increased compared with the respective control (ANOVA, Fisher LSD method). D : A2780WT cells were exposed to isotonic or hypertonic NaCl for 4.5 h in the absence or presence of 100 μM NS3728 or 400 μM DIDS. Data represent 3 individual experiments ± SE. *Significantly increased compared with isotonic control (ANOVA, Fisher LSD method).

Techniques Used: Activation Assay, Caspase-3 Activity Assay

Impairment of swelling induced taurine release following pharmacological inhibition or transient knock-down of LRRC8A in wild-type A2780 cells. Volume-sensitive taurine release and LRRC8A protein expression were measured in ovarian A2780WT cells by tracer technique and Western blot analysis, respectively. A2780WT cells were loaded with [ 3 H]taurine for 2 h, washed, and exposed to isosmotic NaCl medium (300 mOsM) for 10 min and subsequently to hyposmotic medium (200 mOsM) for 20 min (arrow in A indicates shift to hypotonicity). Samples were taken every second minute. A : fractional rate constant (min −1 ) for taurine release was determined and plotted vs. time (min) under isosmotic and hyposmotic conditions in the absence (○) or presence of the VRAC/VSOAC blockers NS3728 (●) or DIDS (■). The anion blockers NS3728 and DIDS were used in the concentration of 10 μM (≈2.5 μM free) and 100 μM, respectively, and present during the isotonic/hypotonic release experiment. Data represent 1 of 3 sets of experiments. B : maximal rate constant for taurine release was determined 6 min after hyposmotic exposure as illustrated in A . Data represent the mean maximal rate constants determined from 3 sets of experiments ± SE. *Significantly reduced compared with control (Student's t -test). C : efficiency of siRNA-mediated LRRC8A knock-down in A2780WT cells was determined by Western blot analysis using control, scramble siRNA and LRRC8A siRNA transfected cells and specific monoclonal antibody raised against human LRRC8A and β-actin (loading control). D : LRRC8A/β-actin protein band intensity ratios were calculated from Western blots shown in C and values given relative to control cells. Data represent mean of 4 experiments ± SE. * and #: Significantly reduced compared with Control and Scramble siRNA, respectively (Student's t -test). Scramble siRNA was not significantly larger than Control ( P = 0.15, Student's t -test). E : maximal rate constant for taurine release was determined at time 6 min after hypotonic exposure in A2780WT (open bar) cells or in A2780 cells exposed to scramble siRNA (light gray bar) or siRNA directed against human LRRC8A (dark gray bar). Data represent mean of 4 experiments ± SE. *Significantly reduced compared with Control (Student's t -test).
Figure Legend Snippet: Impairment of swelling induced taurine release following pharmacological inhibition or transient knock-down of LRRC8A in wild-type A2780 cells. Volume-sensitive taurine release and LRRC8A protein expression were measured in ovarian A2780WT cells by tracer technique and Western blot analysis, respectively. A2780WT cells were loaded with [ 3 H]taurine for 2 h, washed, and exposed to isosmotic NaCl medium (300 mOsM) for 10 min and subsequently to hyposmotic medium (200 mOsM) for 20 min (arrow in A indicates shift to hypotonicity). Samples were taken every second minute. A : fractional rate constant (min −1 ) for taurine release was determined and plotted vs. time (min) under isosmotic and hyposmotic conditions in the absence (○) or presence of the VRAC/VSOAC blockers NS3728 (●) or DIDS (■). The anion blockers NS3728 and DIDS were used in the concentration of 10 μM (≈2.5 μM free) and 100 μM, respectively, and present during the isotonic/hypotonic release experiment. Data represent 1 of 3 sets of experiments. B : maximal rate constant for taurine release was determined 6 min after hyposmotic exposure as illustrated in A . Data represent the mean maximal rate constants determined from 3 sets of experiments ± SE. *Significantly reduced compared with control (Student's t -test). C : efficiency of siRNA-mediated LRRC8A knock-down in A2780WT cells was determined by Western blot analysis using control, scramble siRNA and LRRC8A siRNA transfected cells and specific monoclonal antibody raised against human LRRC8A and β-actin (loading control). D : LRRC8A/β-actin protein band intensity ratios were calculated from Western blots shown in C and values given relative to control cells. Data represent mean of 4 experiments ± SE. * and #: Significantly reduced compared with Control and Scramble siRNA, respectively (Student's t -test). Scramble siRNA was not significantly larger than Control ( P = 0.15, Student's t -test). E : maximal rate constant for taurine release was determined at time 6 min after hypotonic exposure in A2780WT (open bar) cells or in A2780 cells exposed to scramble siRNA (light gray bar) or siRNA directed against human LRRC8A (dark gray bar). Data represent mean of 4 experiments ± SE. *Significantly reduced compared with Control (Student's t -test).

Techniques Used: Inhibition, Expressing, Western Blot, Concentration Assay, Transfection

Administration of anion- and cation-channel blockers reduces Cisplatin-induced expression of p53 and p21 Waf1/Cip1 in wild-type A2780 cells (A2780WT) to the same level as observed in Cisplatin-resistant A2780 cells (A2780CisR). Whole cell protein lysates for Western blot analysis were obtained from A2780WT and A2780CisR cells exposed to 10 μM Cisplatin for 18 h in combination with the anion/amino acid channel blockers NS3728 (100 μM)/DIDS (400 μM) or the cation-channel (TASK-2) blocker clofilium (25 μM). Only A2780WT cells were exposed to channel blockers. Protein expression of LRRC8A, p53, p-p53 (Ser15), p21, and Bax was determined using specific antibodies (see experimental procedures ). β-Actin was used as loading control. A : representative Western blot. B : p53, pp53, p21, and Bax protein expression relative to β-actin in A2780WT and A2780CisR (open bars) control cells and following Cisplatin exposure (black bars). Data represent 4–9 individual sets of experiments where ratios are given relative to the untreated WT cells ± SE. * and #: Expression is significantly increased in Cisplatin-treated cell compared with control cells and significantly reduced in Cisplatin-treated A2780CisR cells compared with Cisplatin-treated A2780WT cells, respectively (ANOVA, Fisher LSD method). C : graph represents the relative effect of Cisplatin (Cisplatin-treated/respective untreated control) on LRRC8A, p53, p-p53, and p21 protein expression in A2780WT, A2780CisR (CisR), and A2780WT cells cotreated with the anion- and cation-channel blockers. Data represent mean of 4–9 individual experiments ± SE. *Significantly reduced effect of Cisplatin compared with A2780WT (ANOVA, Fisher LSD method).
Figure Legend Snippet: Administration of anion- and cation-channel blockers reduces Cisplatin-induced expression of p53 and p21 Waf1/Cip1 in wild-type A2780 cells (A2780WT) to the same level as observed in Cisplatin-resistant A2780 cells (A2780CisR). Whole cell protein lysates for Western blot analysis were obtained from A2780WT and A2780CisR cells exposed to 10 μM Cisplatin for 18 h in combination with the anion/amino acid channel blockers NS3728 (100 μM)/DIDS (400 μM) or the cation-channel (TASK-2) blocker clofilium (25 μM). Only A2780WT cells were exposed to channel blockers. Protein expression of LRRC8A, p53, p-p53 (Ser15), p21, and Bax was determined using specific antibodies (see experimental procedures ). β-Actin was used as loading control. A : representative Western blot. B : p53, pp53, p21, and Bax protein expression relative to β-actin in A2780WT and A2780CisR (open bars) control cells and following Cisplatin exposure (black bars). Data represent 4–9 individual sets of experiments where ratios are given relative to the untreated WT cells ± SE. * and #: Expression is significantly increased in Cisplatin-treated cell compared with control cells and significantly reduced in Cisplatin-treated A2780CisR cells compared with Cisplatin-treated A2780WT cells, respectively (ANOVA, Fisher LSD method). C : graph represents the relative effect of Cisplatin (Cisplatin-treated/respective untreated control) on LRRC8A, p53, p-p53, and p21 protein expression in A2780WT, A2780CisR (CisR), and A2780WT cells cotreated with the anion- and cation-channel blockers. Data represent mean of 4–9 individual experiments ± SE. *Significantly reduced effect of Cisplatin compared with A2780WT (ANOVA, Fisher LSD method).

Techniques Used: Expressing, Western Blot

15) Product Images from "DIDS (4,4'-Diisothiocyanatostilbene-2,2'-disulfonate) directly inhibits caspase activity in HeLa cell lysates"

Article Title: DIDS (4,4'-Diisothiocyanatostilbene-2,2'-disulfonate) directly inhibits caspase activity in HeLa cell lysates

Journal: Cell Death Discovery

doi: 10.1038/cddiscovery.2015.37

DIDS totally eliminates caspase activities in a cell-free extract. HeLa cells that were previously treated with or without STS (1 μ M) for 4 h were lysed and these extracts were incubated with DIDS (50 nM), 2-APB (50 nM) or flufenamic acid (50 nM) for 45 min at 37 °C followed by the corresponding caspase activity procedure. Note that DIDS but not the other ion channel inhibitors abolished caspase-3 ( a , n =5), caspase-9 (LEHDase) and caspase-8 (lETDase) activities ( b , n =3). Neither 2-APB nor flufenamic acid induced any DEVDase activity (not shown). * P
Figure Legend Snippet: DIDS totally eliminates caspase activities in a cell-free extract. HeLa cells that were previously treated with or without STS (1 μ M) for 4 h were lysed and these extracts were incubated with DIDS (50 nM), 2-APB (50 nM) or flufenamic acid (50 nM) for 45 min at 37 °C followed by the corresponding caspase activity procedure. Note that DIDS but not the other ion channel inhibitors abolished caspase-3 ( a , n =5), caspase-9 (LEHDase) and caspase-8 (lETDase) activities ( b , n =3). Neither 2-APB nor flufenamic acid induced any DEVDase activity (not shown). * P

Techniques Used: Incubation, Activity Assay

DIDS inhibits caspase-3 processing, PARP degradation and apoptotic nuclei features. HeLa cells preincubated with or without DIDS (50 or 500 μ M) for 30 min before incubation with STS (1 μ M) for 4 h were lysed and ( a ) pro-caspase 3 was detected with antibody (clone 3G2, n =3) and ( b ) PARP fragmentation with antibody (clone 4C10-5, n =5). DIDS totally abolished procaspase-3 proteolytic processing and strongly diminished STS-induced PARP fragmentation. ( c ) Apoptotic nuclei were induced by STS (4 h incubation). However, neither the vehicle (DMSO, 0.1 or 1%) nor DIDS alone induced apoptotic nuclei. Preincubation with DIDS (50 μ M) strongly inhibited STS-induced apoptotic nuclei features. Interestingly, this inhibitory effect was a little bit smaller for a 10 times higher concentration of DIDS (500 μ M) * , P
Figure Legend Snippet: DIDS inhibits caspase-3 processing, PARP degradation and apoptotic nuclei features. HeLa cells preincubated with or without DIDS (50 or 500 μ M) for 30 min before incubation with STS (1 μ M) for 4 h were lysed and ( a ) pro-caspase 3 was detected with antibody (clone 3G2, n =3) and ( b ) PARP fragmentation with antibody (clone 4C10-5, n =5). DIDS totally abolished procaspase-3 proteolytic processing and strongly diminished STS-induced PARP fragmentation. ( c ) Apoptotic nuclei were induced by STS (4 h incubation). However, neither the vehicle (DMSO, 0.1 or 1%) nor DIDS alone induced apoptotic nuclei. Preincubation with DIDS (50 μ M) strongly inhibited STS-induced apoptotic nuclei features. Interestingly, this inhibitory effect was a little bit smaller for a 10 times higher concentration of DIDS (500 μ M) * , P

Techniques Used: Incubation, Concentration Assay

DIDS inhibits STS-induced caspase activation in HeLa cells. Cells that were preincubated with or without DIDS (50 or 500 μ M) for 30 min were incubated with STS 1 μ M for 4 h and lysed to determine caspase activity, which was normalized based on the response obtained with STS. Panels show caspase-3 ( a , n =7), capase-9 ( b , n =4) and caspase-8 ( c , n =4) activities without (total activity, solid columns) and with caspase inhibitors (open columns), which were Ac-DEVD-CHO for caspase-3 ( a ), Ac-LEHD-CHO for caspase-9 ( b ) and Ac-IETD-CHO for capase-8 ( c ).ln all cases, STS induced a significant caspase activity with respect to control (** P
Figure Legend Snippet: DIDS inhibits STS-induced caspase activation in HeLa cells. Cells that were preincubated with or without DIDS (50 or 500 μ M) for 30 min were incubated with STS 1 μ M for 4 h and lysed to determine caspase activity, which was normalized based on the response obtained with STS. Panels show caspase-3 ( a , n =7), capase-9 ( b , n =4) and caspase-8 ( c , n =4) activities without (total activity, solid columns) and with caspase inhibitors (open columns), which were Ac-DEVD-CHO for caspase-3 ( a ), Ac-LEHD-CHO for caspase-9 ( b ) and Ac-IETD-CHO for capase-8 ( c ).ln all cases, STS induced a significant caspase activity with respect to control (** P

Techniques Used: Activation Assay, Incubation, Activity Assay

DIDS does not abolish STS-induced cyt c release. HeLa cells that were in the absence of serum for 19.5 h were preincubated with or without DIDS (50 or 500 μ M) for 30 min followed by STS (1 μ M) incubation for 4 h and lysed to determine cytochrome c release from mitochondria. ( a ) Representative western blots of cytochrome c (cyt c) and p tubulin in supernatants of cell lysates. Note that DIDS 500 μ M release some cyt c by itself. ( b ) The optical density ratio between signals of cyt c and p-tubulin was normalized using the STS-induced ratio as 100%. DIDS did not inhibit cytochrome c release, only reduced it by 20% at both concentrations of DIDS ( n =7). * P
Figure Legend Snippet: DIDS does not abolish STS-induced cyt c release. HeLa cells that were in the absence of serum for 19.5 h were preincubated with or without DIDS (50 or 500 μ M) for 30 min followed by STS (1 μ M) incubation for 4 h and lysed to determine cytochrome c release from mitochondria. ( a ) Representative western blots of cytochrome c (cyt c) and p tubulin in supernatants of cell lysates. Note that DIDS 500 μ M release some cyt c by itself. ( b ) The optical density ratio between signals of cyt c and p-tubulin was normalized using the STS-induced ratio as 100%. DIDS did not inhibit cytochrome c release, only reduced it by 20% at both concentrations of DIDS ( n =7). * P

Techniques Used: Incubation, Western Blot

16) Product Images from "Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia"

Article Title: Chloride channels are involved in sperm motility and are downregulated in spermatozoa from patients with asthenozoospermia

Journal: Asian Journal of Andrology

doi: 10.4103/1008-682X.181816

Inhibitory effects of the chloride channel blockers NPPB and DIDS (both at 100 μmol l −1 ) on the motility of spermatozoa from the normozoospermic and asthenozoospermic semen samples in hypotonic BWW 290 . The inhibitory effects of NPPB and DIDS on the (a) progressive motility (PR), (b) straight-line velocity (VSL), (c) curved-line velocity (VCL) and (d) average path velocity (VAP), respectively, when incubated for 30 min. Data are shown as the mean ± s.e. (semen samples, n from 8 to 22). ** P
Figure Legend Snippet: Inhibitory effects of the chloride channel blockers NPPB and DIDS (both at 100 μmol l −1 ) on the motility of spermatozoa from the normozoospermic and asthenozoospermic semen samples in hypotonic BWW 290 . The inhibitory effects of NPPB and DIDS on the (a) progressive motility (PR), (b) straight-line velocity (VSL), (c) curved-line velocity (VCL) and (d) average path velocity (VAP), respectively, when incubated for 30 min. Data are shown as the mean ± s.e. (semen samples, n from 8 to 22). ** P

Techniques Used: Incubation

Effects of hypotonic stimulation and the chloride channel blockers NPPB and DIDS on the volumes of spermatozoa from normozoospermic and asthenozoospermic semen samples. Sperm volume was detected by monitoring the forward scatter signals (FSCs) using flow cytometry. (a) Relative sperm volumes expressed as FSC signals of the in the hypotonic (BWW 290 ) and isotonic (BWW 330 ) solutions; (b) and (c) the effects of NPPB and DIDS (both at 100 μmol l −1 ) on the relative sperm volume (expressed as the FSC signal ratio of blockers/BWW 290 control) in the normozoospermic and asthenozoospermic semen samples, respectively; and the percentage sperm volume increases caused by (d) DIDS and (e) NPPB in the hypotonic solution, respectively. ** P
Figure Legend Snippet: Effects of hypotonic stimulation and the chloride channel blockers NPPB and DIDS on the volumes of spermatozoa from normozoospermic and asthenozoospermic semen samples. Sperm volume was detected by monitoring the forward scatter signals (FSCs) using flow cytometry. (a) Relative sperm volumes expressed as FSC signals of the in the hypotonic (BWW 290 ) and isotonic (BWW 330 ) solutions; (b) and (c) the effects of NPPB and DIDS (both at 100 μmol l −1 ) on the relative sperm volume (expressed as the FSC signal ratio of blockers/BWW 290 control) in the normozoospermic and asthenozoospermic semen samples, respectively; and the percentage sperm volume increases caused by (d) DIDS and (e) NPPB in the hypotonic solution, respectively. ** P

Techniques Used: Flow Cytometry, Cytometry

17) Product Images from "Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism"

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism

Journal: Scientific Reports

doi: 10.1038/srep31433

Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p
Figure Legend Snippet: Effects of DIDS (400 μM) ( A ), AZ (200 μM) ( B ), EIPA (400 μM) ( C ), and bumetanide (Bumex; 400 μM) and/or DIDS (400 μM) ( D ) on Cl − flux at ionocytes of larval skin. Data are presented as the means ± SEM. * p

Techniques Used:

18) Product Images from "The nuclear chloride ion channel NCC27 is involved in regulation of the cell cycle"

Article Title: The nuclear chloride ion channel NCC27 is involved in regulation of the cell cycle

Journal: The Journal of Physiology

doi: 10.1111/j.1469-7793.2000.00541.x

Effects of chloride ion channel blockers on single-channel NCC27 currents The top panel demonstrates the effect of IAA-94. Current blockade begins within 30 s and is complete at 3 min, but recovers fully and rapidly. A9C (middle panel) produces rapid, complete but irreversible block. DIDS had no effect on the NCC27 single-channel current (bottom panel).
Figure Legend Snippet: Effects of chloride ion channel blockers on single-channel NCC27 currents The top panel demonstrates the effect of IAA-94. Current blockade begins within 30 s and is complete at 3 min, but recovers fully and rapidly. A9C (middle panel) produces rapid, complete but irreversible block. DIDS had no effect on the NCC27 single-channel current (bottom panel).

Techniques Used: Blocking Assay

Line graphs from 3 independent experiments demonstrating the effects of increasing concentrations of Cl − channel blockers on the proportion of CHO-K1 cells in G2/M phase (expressed as % change from control) DIDS had no effect at any concentration (left). A9C significantly increased the fraction in G2/M at 2 mM ( P
Figure Legend Snippet: Line graphs from 3 independent experiments demonstrating the effects of increasing concentrations of Cl − channel blockers on the proportion of CHO-K1 cells in G2/M phase (expressed as % change from control) DIDS had no effect at any concentration (left). A9C significantly increased the fraction in G2/M at 2 mM ( P

Techniques Used: Concentration Assay

19) Product Images from "Patch Clamp on the Luminal Membrane of Exocrine Gland Acini from Frog Skin (Rana esculenta) Reveals the Presence of Cystic Fibrosis Transmembrane Conductance Regulator-like Cl− Channels Activated by Cyclic AMP "

Article Title: Patch Clamp on the Luminal Membrane of Exocrine Gland Acini from Frog Skin (Rana esculenta) Reveals the Presence of Cystic Fibrosis Transmembrane Conductance Regulator-like Cl− Channels Activated by Cyclic AMP

Journal: The Journal of General Physiology

doi:

Pharmacology. Inside-out patch with two active Cl − channels held at −V p = 50 mV. In “Control” periods 1–5, the patch was washed with I-28 + 1.5 mM ATP on the inside. Sequential addition of putative blockers to I-28 + 1.5 mM ATP are indicated. In Control period 5, the patch was washed for 6 min without DIDS. Note the appearance of channel transitions with depressed amplitude.
Figure Legend Snippet: Pharmacology. Inside-out patch with two active Cl − channels held at −V p = 50 mV. In “Control” periods 1–5, the patch was washed with I-28 + 1.5 mM ATP on the inside. Sequential addition of putative blockers to I-28 + 1.5 mM ATP are indicated. In Control period 5, the patch was washed for 6 min without DIDS. Note the appearance of channel transitions with depressed amplitude.

Techniques Used:

20) Product Images from "Cl− channel is required for CXCL10-induced neuronal activation and itch response in a murine model of allergic contact dermatitis"

Article Title: Cl− channel is required for CXCL10-induced neuronal activation and itch response in a murine model of allergic contact dermatitis

Journal: Journal of Neurophysiology

doi: 10.1152/jn.00187.2017

Effects of Cl − channel antagonists and intracellular Ca 2+ on CXCL10-induced currents in DRG neurons innervating CHS skin. A and B : sample traces of the I CXCL10 recorded in the absence ( A ) and presence of the Cl − channel blockers DIDS (100 µM; B ) or NFA (100 µM; C ) applied to the bath or in the presence of 10 mM BAPTA in the pipette solution ( D ). The high-[Cl − ] i pipette solution was used. E : pretreatment with DIDS or NFA for 3 min significantly reduced the peak amplitude of I CXCL10 . Replacement of 11 mM EGTA with 10 mM BAPTA in the pipette solution almost abolished this inward current. The numbers of DRG neurons tested are given in parentheses. * P
Figure Legend Snippet: Effects of Cl − channel antagonists and intracellular Ca 2+ on CXCL10-induced currents in DRG neurons innervating CHS skin. A and B : sample traces of the I CXCL10 recorded in the absence ( A ) and presence of the Cl − channel blockers DIDS (100 µM; B ) or NFA (100 µM; C ) applied to the bath or in the presence of 10 mM BAPTA in the pipette solution ( D ). The high-[Cl − ] i pipette solution was used. E : pretreatment with DIDS or NFA for 3 min significantly reduced the peak amplitude of I CXCL10 . Replacement of 11 mM EGTA with 10 mM BAPTA in the pipette solution almost abolished this inward current. The numbers of DRG neurons tested are given in parentheses. * P

Techniques Used: Transferring

21) Product Images from "Euryhaline pufferfish NBCe1 differs from nonmarine species NBCe1 physiology"

Article Title: Euryhaline pufferfish NBCe1 differs from nonmarine species NBCe1 physiology

Journal: American Journal of Physiology - Cell Physiology

doi: 10.1152/ajpcell.00233.2011

Experimental protocol used to study HCO 3 − -independent components and DIDS inhibition of NBCe1 proteins. A and B : current traces ( V h = −60 mV, n = 3) from fNBCe1 and hkNBCe1. Solution changes are indicated by horizontalbars above traces.
Figure Legend Snippet: Experimental protocol used to study HCO 3 − -independent components and DIDS inhibition of NBCe1 proteins. A and B : current traces ( V h = −60 mV, n = 3) from fNBCe1 and hkNBCe1. Solution changes are indicated by horizontalbars above traces.

Techniques Used: Inhibition

DIDS effects on CO 2 /HCO 3 − -elicited currents of NBCe1 proteins. A and B : DIDSinhibition of CO 2 /HCO 3 − -elicited currents in oocytes expressing fNBCe1 and hkNBCe1. Experimental protocolsare described in legend. I-V relationships show
Figure Legend Snippet: DIDS effects on CO 2 /HCO 3 − -elicited currents of NBCe1 proteins. A and B : DIDSinhibition of CO 2 /HCO 3 − -elicited currents in oocytes expressing fNBCe1 and hkNBCe1. Experimental protocolsare described in legend. I-V relationships show

Techniques Used: Expressing

22) Product Images from "Assessment of Methods for the Intracellular Blockade of GABAA Receptors"

Article Title: Assessment of Methods for the Intracellular Blockade of GABAA Receptors

Journal: PLoS ONE

doi: 10.1371/journal.pone.0160900

Effect of DIDS, the absence of nucleotides and KF on evoked IPSCs. Upper panels of A-D: Example traces during the diffusion of three intracellular pipette solutions (K-Gluconate +DIDS (A), K-Gluconate without ATP or GTP (B), and K-Fluoride at -80mV (C) and -50mV (D)). Each trace shows the IPSC at the start of recording (black, time point i in middle panels), in the last minute before bath application of picrotoxin (magenta, time point ii in middle panels) and at the end of the bath application of picrotoxin (blue, time point iii in middle panels). Middle panels of A-D: Normalised IPSC amplitudes. Blue bar indicates the presence of bath picrotoxin. Lower panels of A-D: Corresponding series resistance during experiments. NBQX and AP5 were present throughout the experiments. E) Group data of the normalised IPSC amplitudes, taken at time point ii, individual data points marked with magenta dots. Summary statistics represent tests comparing the normalised evoked IPSC amplitudes at time point ii with the normalised baseline amplitude for each data set. Data are plotted as mean ± SEM.
Figure Legend Snippet: Effect of DIDS, the absence of nucleotides and KF on evoked IPSCs. Upper panels of A-D: Example traces during the diffusion of three intracellular pipette solutions (K-Gluconate +DIDS (A), K-Gluconate without ATP or GTP (B), and K-Fluoride at -80mV (C) and -50mV (D)). Each trace shows the IPSC at the start of recording (black, time point i in middle panels), in the last minute before bath application of picrotoxin (magenta, time point ii in middle panels) and at the end of the bath application of picrotoxin (blue, time point iii in middle panels). Middle panels of A-D: Normalised IPSC amplitudes. Blue bar indicates the presence of bath picrotoxin. Lower panels of A-D: Corresponding series resistance during experiments. NBQX and AP5 were present throughout the experiments. E) Group data of the normalised IPSC amplitudes, taken at time point ii, individual data points marked with magenta dots. Summary statistics represent tests comparing the normalised evoked IPSC amplitudes at time point ii with the normalised baseline amplitude for each data set. Data are plotted as mean ± SEM.

Techniques Used: Diffusion-based Assay, Transferring

Effect of DNDS, picrotoxin and CsF-DIDS on evoked IPSCs. Upper panels of A-E: Example traces during the diffusion of K-Gluconate (A), K-Gluconate +DNDS (B), K-Gluconate +DNDS CaCl 2 (C), K-Gluconate + picrotoxin (D) and Cs-Fluoride +DIDS (E) intracellular pipette solutions. Each trace shows the IPSC at the start of recording (black, time point i in middle panels), in the last minute before bath application of picrotoxin (magenta, time point ii in middle panels) and at the end of the bath application of picrotoxin (blue, time point iii in middle panels). Middle panels of A-E: Normalised IPSC amplitudes. Blue bar indicates the presence of bath picrotoxin (PTX). Lower panels of A-D: Corresponding series resistance during experiments. NBQX and AP5 were present throughout the experiments. E) Group data of the normalised evoked IPSC amplitudes for the four intracellular pipette solutions, taken at time point ii, individual data points marked as magenta dots. Summary statistics represent tests comparing the normalised evoked IPSC amplitudes at time point ii with the normalised baseline amplitude for each data set. Data are plotted as mean ± SEM.
Figure Legend Snippet: Effect of DNDS, picrotoxin and CsF-DIDS on evoked IPSCs. Upper panels of A-E: Example traces during the diffusion of K-Gluconate (A), K-Gluconate +DNDS (B), K-Gluconate +DNDS CaCl 2 (C), K-Gluconate + picrotoxin (D) and Cs-Fluoride +DIDS (E) intracellular pipette solutions. Each trace shows the IPSC at the start of recording (black, time point i in middle panels), in the last minute before bath application of picrotoxin (magenta, time point ii in middle panels) and at the end of the bath application of picrotoxin (blue, time point iii in middle panels). Middle panels of A-E: Normalised IPSC amplitudes. Blue bar indicates the presence of bath picrotoxin (PTX). Lower panels of A-D: Corresponding series resistance during experiments. NBQX and AP5 were present throughout the experiments. E) Group data of the normalised evoked IPSC amplitudes for the four intracellular pipette solutions, taken at time point ii, individual data points marked as magenta dots. Summary statistics represent tests comparing the normalised evoked IPSC amplitudes at time point ii with the normalised baseline amplitude for each data set. Data are plotted as mean ± SEM.

Techniques Used: Diffusion-based Assay, Transferring

23) Product Images from "DUAL AND OPPOSITE EFFECTS OF hRAD51 CHEMICAL MODULATION ON HIV-1 INTEGRATION"

Article Title: DUAL AND OPPOSITE EFFECTS OF hRAD51 CHEMICAL MODULATION ON HIV-1 INTEGRATION

Journal: Chemistry & biology

doi: 10.1016/j.chembiol.2015.04.020

Effect of hRAD51 modulators on HIV-1 integration The chemical structure of the stimulatory compounds RS-1 and P-ter ( A ) and the inhibitory compounds RI-1 and DIDS, as well as the sequence of the aptamers ( B ) are indicated. Increasing concentrations of wt-hRAD51 were added in a standard concerted integration assay in the presence of 100 µM ATP (w/o molecule or aptamer), and with 7.5 or 15 µM RS-1 ( C ), P-ter ( D ), RI-1 ( E ) or 0.1, 0.25 or 0.5 µM of A30 ( F , was normalized to 100 %.
Figure Legend Snippet: Effect of hRAD51 modulators on HIV-1 integration The chemical structure of the stimulatory compounds RS-1 and P-ter ( A ) and the inhibitory compounds RI-1 and DIDS, as well as the sequence of the aptamers ( B ) are indicated. Increasing concentrations of wt-hRAD51 were added in a standard concerted integration assay in the presence of 100 µM ATP (w/o molecule or aptamer), and with 7.5 or 15 µM RS-1 ( C ), P-ter ( D ), RI-1 ( E ) or 0.1, 0.25 or 0.5 µM of A30 ( F , was normalized to 100 %.

Techniques Used: Sequencing

24) Product Images from "DIDS inhibits Na-K-ATPase activity in porcine nonpigmented ciliary epithelial cells by a Src family kinase-dependent mechanism"

Article Title: DIDS inhibits Na-K-ATPase activity in porcine nonpigmented ciliary epithelial cells by a Src family kinase-dependent mechanism

Journal: American Journal of Physiology - Cell Physiology

doi: 10.1152/ajpcell.00057.2013

The influence of DIDS on ERK1/2 phosphorylation. A : a typical time course study in cells exposed to DIDS (100 μM). B : effect of U0126 (10 μM) on DIDS-induced ERK1/2 phosphorylation. Cells were preincubated with U0126 (10 μM) for
Figure Legend Snippet: The influence of DIDS on ERK1/2 phosphorylation. A : a typical time course study in cells exposed to DIDS (100 μM). B : effect of U0126 (10 μM) on DIDS-induced ERK1/2 phosphorylation. Cells were preincubated with U0126 (10 μM) for

Techniques Used:

The influence of U0126 on Na-K-ATPase activity in cells exposed to DIDS. Na-K-ATPase activity (ouabain-sensitive ATP hydrolysis) was measured in samples obtained by homogenizing cells that had been preincubated with U0126 (10 μM) for 20 min before
Figure Legend Snippet: The influence of U0126 on Na-K-ATPase activity in cells exposed to DIDS. Na-K-ATPase activity (ouabain-sensitive ATP hydrolysis) was measured in samples obtained by homogenizing cells that had been preincubated with U0126 (10 μM) for 20 min before

Techniques Used: Activity Assay

Comparison of Na-K-ATPase activity inhibition elicited by DIDS and SITS. Na-K-ATPase activity (ouabain-sensitive ATP hydrolysis) was measured in samples obtained by homogenizing cells that had been exposed to DIDS (100 μM) or SITS (100 μM)
Figure Legend Snippet: Comparison of Na-K-ATPase activity inhibition elicited by DIDS and SITS. Na-K-ATPase activity (ouabain-sensitive ATP hydrolysis) was measured in samples obtained by homogenizing cells that had been exposed to DIDS (100 μM) or SITS (100 μM)

Techniques Used: Activity Assay, Inhibition

25) Product Images from "Interaction of stilbene disulphonates with cloned KATP channels"

Article Title: Interaction of stilbene disulphonates with cloned KATP channels

Journal: British Journal of Pharmacology

doi: 10.1038/sj.bjp.0703916

Effects of stilbene disulphonates on Kir6.2ΔC-C42A/SUR1 and Kir6.2ΔC-K67Q/SUR1 currents. Macroscopic Kir6.2ΔC-C42A/SUR1 (A) or Kir6.2ΔC-K67Q/SUR1 (B, C) current recorded at −60 mV in an inside-out patch. ATP, SITS or DIDS were added as indicated by the bars. The dashed line indicates the zero current level.
Figure Legend Snippet: Effects of stilbene disulphonates on Kir6.2ΔC-C42A/SUR1 and Kir6.2ΔC-K67Q/SUR1 currents. Macroscopic Kir6.2ΔC-C42A/SUR1 (A) or Kir6.2ΔC-K67Q/SUR1 (B, C) current recorded at −60 mV in an inside-out patch. ATP, SITS or DIDS were added as indicated by the bars. The dashed line indicates the zero current level.

Techniques Used:

26) Product Images from "Abscisic Acid Transport in Human Erythrocytes *"

Article Title: Abscisic Acid Transport in Human Erythrocytes *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M114.629501

The ABA antagonist #10 and Band 3 inhibitor DIDS inhibit the ABA-induced [cAMP] i rise and ATP release. A, RBC were preincubated at a 50% hematocrit with 250 μ m of the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine for 15 min; cells
Figure Legend Snippet: The ABA antagonist #10 and Band 3 inhibitor DIDS inhibit the ABA-induced [cAMP] i rise and ATP release. A, RBC were preincubated at a 50% hematocrit with 250 μ m of the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine for 15 min; cells

Techniques Used:

27) Product Images from "In Mycoplasma hominis the OppA-mediated cytoadhesion depends on its ATPase activity"

Article Title: In Mycoplasma hominis the OppA-mediated cytoadhesion depends on its ATPase activity

Journal: BMC Microbiology

doi: 10.1186/1471-2180-11-185

Adherence of OppA to HeLa cells in the presence of ATPase inhibitors . OppA (black bars) or P60/P80 as a control (white bars), (0.5 μg OppA/well and 0.3 μg P60/well) were preincubated with 200μM DIDS, suramin, ouabain or oligomycin for 20 min before analyzing in adhesion assay. ATPase activity (A) and adhesion efficiency (B) were measured and depicted in relation to the untreated OppA. OppA (0.5 μg protein) was preincubated with FSBA or MgATP for 20 min and then added to HeLa cells (C). Adherence of OppA to HeLa cells in dependence on supplement concentration was determined as described in Material and Methods. Data represent means of three independent experiments with triplicate samples in each experiment. Statistical analysis was performed by unpaired t-test and statistically significant results designated by *. *P
Figure Legend Snippet: Adherence of OppA to HeLa cells in the presence of ATPase inhibitors . OppA (black bars) or P60/P80 as a control (white bars), (0.5 μg OppA/well and 0.3 μg P60/well) were preincubated with 200μM DIDS, suramin, ouabain or oligomycin for 20 min before analyzing in adhesion assay. ATPase activity (A) and adhesion efficiency (B) were measured and depicted in relation to the untreated OppA. OppA (0.5 μg protein) was preincubated with FSBA or MgATP for 20 min and then added to HeLa cells (C). Adherence of OppA to HeLa cells in dependence on supplement concentration was determined as described in Material and Methods. Data represent means of three independent experiments with triplicate samples in each experiment. Statistical analysis was performed by unpaired t-test and statistically significant results designated by *. *P

Techniques Used: Cell Adhesion Assay, Activity Assay, Concentration Assay

28) Product Images from "Spontaneous electrical rhythmicity in cultured interstitial cells of Cajal from the murine small intestine"

Article Title: Spontaneous electrical rhythmicity in cultured interstitial cells of Cajal from the murine small intestine

Journal: The Journal of Physiology

doi: 10.1111/j.1469-7793.1998.203by.x

Pharmacology of slow wave oscillations in cultured ICC A , lack of effect of nisoldipine (1 μm) on slow waves. B , inhibition of slow waves when cells were exposed to a bathing solution in which Ca 2+ was replaced by Mn 2+ (in physiological saline solution, MnPSS). C , effects of cyclopiazonic acid (CPA). CPA increased frequency and produced a small hyperpolarization in MDP. D , gadolinium (Gd 3+ ) blocked slow wave activity. E , DIDS reduced the amplitude of slow waves, caused a small hyperpolarization in MDP, but did not block spontaneous rhythmicity.
Figure Legend Snippet: Pharmacology of slow wave oscillations in cultured ICC A , lack of effect of nisoldipine (1 μm) on slow waves. B , inhibition of slow waves when cells were exposed to a bathing solution in which Ca 2+ was replaced by Mn 2+ (in physiological saline solution, MnPSS). C , effects of cyclopiazonic acid (CPA). CPA increased frequency and produced a small hyperpolarization in MDP. D , gadolinium (Gd 3+ ) blocked slow wave activity. E , DIDS reduced the amplitude of slow waves, caused a small hyperpolarization in MDP, but did not block spontaneous rhythmicity.

Techniques Used: Cell Culture, Immunocytochemistry, Inhibition, Produced, Activity Assay, Blocking Assay

29) Product Images from "Neuronal and Astroglial Correlates Underlying Spatiotemporal Intrinsic Optical Signal in the Rat Hippocampal Slice"

Article Title: Neuronal and Astroglial Correlates Underlying Spatiotemporal Intrinsic Optical Signal in the Rat Hippocampal Slice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0057694

Neuronal K + /Cl − cotransporter, non-specific Cl − and volume-regulated anion channels contribute to IOS. A Left and Middle: Representative IOS amplitude map and field response curve under control condition and 5 mM furosemide application, respectively. The colorbar indicates the maximum change of the transmittance compared to the resting light intensity. A Right: Spatial visualization of the percentage of control changes of IOS signal caused by furosemide application. B Left and Middle: Representative IOS amplitude map and field response curve under control condition and 200 µM DIDS application, respectively. The colorbar indicates the maximum change of the transmittance compared to the resting light intensity. B Right: Spatial visualization of the percentage of control changes of IOS signal caused by DIDS application. C: The effect of 5 mM furosemide, 10 µM bumetanide, 200 µM DIDS and 40 µM DCPIB on the field response and IOS parameters in percentage of control. Asterisks indicate significant changes compared to control (P
Figure Legend Snippet: Neuronal K + /Cl − cotransporter, non-specific Cl − and volume-regulated anion channels contribute to IOS. A Left and Middle: Representative IOS amplitude map and field response curve under control condition and 5 mM furosemide application, respectively. The colorbar indicates the maximum change of the transmittance compared to the resting light intensity. A Right: Spatial visualization of the percentage of control changes of IOS signal caused by furosemide application. B Left and Middle: Representative IOS amplitude map and field response curve under control condition and 200 µM DIDS application, respectively. The colorbar indicates the maximum change of the transmittance compared to the resting light intensity. B Right: Spatial visualization of the percentage of control changes of IOS signal caused by DIDS application. C: The effect of 5 mM furosemide, 10 µM bumetanide, 200 µM DIDS and 40 µM DCPIB on the field response and IOS parameters in percentage of control. Asterisks indicate significant changes compared to control (P

Techniques Used:

30) Product Images from "HIV-1 Tat protein directly induces mitochondrial membrane permeabilization and inactivates cytochrome c oxidase"

Article Title: HIV-1 Tat protein directly induces mitochondrial membrane permeabilization and inactivates cytochrome c oxidase

Journal: Cell Death & Disease

doi: 10.1038/cddis.2012.21

Tat-induced swelling in liver isolated mitochondria. ( a ) Sequence of full-length Tat[1-86] (HIV-1 Lai) and Tat derived peptides. ( b ) Dose/time response of Tat[1-86]-induced swelling. Isolated mouse liver mitochondria were exposed to full-length Tat at the indicated concentrations and mitochondrial swelling (measured as 90° light scattering at 545 nm) was monitored continuously. ( c ) Comparative analysis of the effect of Tat-derived peptides on mitochondrial swelling. Isolated mouse liver mitochondria were exposed to the indicated concentrations of Tat-derived peptides. Mitochondrial swelling was monitored for 30 min. Percentages of mitochondrial swelling were calculated as described under Materials and Methods. Data are means (±S.D.) of three independent experiments. ( d ) Evaluation of PTP-related inhibitors on mitochondrial swelling. Liver mitochondria were exposed to Tat[1-86] (0.3 μ M; 30 min) in the presence or absence (Co.) of the following compounds (added 5 min before Tat): cyclosporin A (CsA; 30 μ M), ADP (1 mM), bongkrekic acid (BA; 50 μ M), Bcl-2 (400 nM), Bcl-XL (400 nM), or DIDS (5 μ M). Histograms represent mean values (±S.D.) of five independent experiments. * P
Figure Legend Snippet: Tat-induced swelling in liver isolated mitochondria. ( a ) Sequence of full-length Tat[1-86] (HIV-1 Lai) and Tat derived peptides. ( b ) Dose/time response of Tat[1-86]-induced swelling. Isolated mouse liver mitochondria were exposed to full-length Tat at the indicated concentrations and mitochondrial swelling (measured as 90° light scattering at 545 nm) was monitored continuously. ( c ) Comparative analysis of the effect of Tat-derived peptides on mitochondrial swelling. Isolated mouse liver mitochondria were exposed to the indicated concentrations of Tat-derived peptides. Mitochondrial swelling was monitored for 30 min. Percentages of mitochondrial swelling were calculated as described under Materials and Methods. Data are means (±S.D.) of three independent experiments. ( d ) Evaluation of PTP-related inhibitors on mitochondrial swelling. Liver mitochondria were exposed to Tat[1-86] (0.3 μ M; 30 min) in the presence or absence (Co.) of the following compounds (added 5 min before Tat): cyclosporin A (CsA; 30 μ M), ADP (1 mM), bongkrekic acid (BA; 50 μ M), Bcl-2 (400 nM), Bcl-XL (400 nM), or DIDS (5 μ M). Histograms represent mean values (±S.D.) of five independent experiments. * P

Techniques Used: Isolation, Sequencing, Derivative Assay

Tat-induced swelling in heart-isolated mitochondria. Isolated mouse heart mitochondria were exposed to full-length Tat[1-86] 0.6 μ M and mitochondrial swelling (measured as 90° light scattering at 545 nm) was monitored for 30 min. When indicated, mitochondria were preexposed for 5 min in the presence or absence of the following compounds: cyclosporin A (CsA; 30 μ M), ADP (1 mM), bongkrekic acid (BA; 50 μ M), Bcl-2 (400 nM), or DIDS (5 μ M). Then, mitochondria were incubated with Tat[1-86] (0.3 μ M; 30 min). Percentages of mitochondrial swelling (left panel) were calculated as described under Materials and Methods. Positive control was defined by the addition of 50 μ M CaCl 2 . Histograms represent mean values (±S.D.) of three independent experiments
Figure Legend Snippet: Tat-induced swelling in heart-isolated mitochondria. Isolated mouse heart mitochondria were exposed to full-length Tat[1-86] 0.6 μ M and mitochondrial swelling (measured as 90° light scattering at 545 nm) was monitored for 30 min. When indicated, mitochondria were preexposed for 5 min in the presence or absence of the following compounds: cyclosporin A (CsA; 30 μ M), ADP (1 mM), bongkrekic acid (BA; 50 μ M), Bcl-2 (400 nM), or DIDS (5 μ M). Then, mitochondria were incubated with Tat[1-86] (0.3 μ M; 30 min). Percentages of mitochondrial swelling (left panel) were calculated as described under Materials and Methods. Positive control was defined by the addition of 50 μ M CaCl 2 . Histograms represent mean values (±S.D.) of three independent experiments

Techniques Used: Isolation, Incubation, Positive Control

31) Product Images from "Depolarization-induced depression of inhibitory transmission in cerebellar Purkinje cells"

Article Title: Depolarization-induced depression of inhibitory transmission in cerebellar Purkinje cells

Journal: Physiological Reports

doi: 10.1002/phy2.61

CaCC blockers disturbed DDI occurrence. (A) The experiments were performed using a pipette filled with NFA (10 μmol/L, n = 7), (B) DIDS (10 μmol/L, n = 8), and (C) vehicle (DMSO; 0.01%, n = 6). The left panel (a) represents the time course of the experiments. The right panel (b) shows representative traces from each experiment. The traces of a (before, at t = 0) and b (after, at t = 10) correspond to the timings shown on the left in the figures. Depolarizing trains were applied at t = 5, arrow. (D) a. Current-voltage relationships of calcium currents in the presence of NFA (filled circles, n = 8) and DMSO (open circles, n = 8). Data obtained from vehicle experiments with DMSO were used as a control. Inset; representative trace of voltage-dependent calcium current was recorded at a holding voltage of −70.5 mV to 20.5 mV with NFA in the pipette. b. Representative traces of chloride tail currents, elicited by holding voltages of −80.5 mV and −60.5 mV. NFA blocked the tail current of CaCCs ( n = 8). Bar graph shows the effect of NFA on the charge of the tail current ( P
Figure Legend Snippet: CaCC blockers disturbed DDI occurrence. (A) The experiments were performed using a pipette filled with NFA (10 μmol/L, n = 7), (B) DIDS (10 μmol/L, n = 8), and (C) vehicle (DMSO; 0.01%, n = 6). The left panel (a) represents the time course of the experiments. The right panel (b) shows representative traces from each experiment. The traces of a (before, at t = 0) and b (after, at t = 10) correspond to the timings shown on the left in the figures. Depolarizing trains were applied at t = 5, arrow. (D) a. Current-voltage relationships of calcium currents in the presence of NFA (filled circles, n = 8) and DMSO (open circles, n = 8). Data obtained from vehicle experiments with DMSO were used as a control. Inset; representative trace of voltage-dependent calcium current was recorded at a holding voltage of −70.5 mV to 20.5 mV with NFA in the pipette. b. Representative traces of chloride tail currents, elicited by holding voltages of −80.5 mV and −60.5 mV. NFA blocked the tail current of CaCCs ( n = 8). Bar graph shows the effect of NFA on the charge of the tail current ( P

Techniques Used: Transferring

32) Product Images from "CFTR-mediated halide transport in phagosomes of human neutrophils"

Article Title: CFTR-mediated halide transport in phagosomes of human neutrophils

Journal: Journal of Leukocyte Biology

doi: 10.1189/jlb.1009655

Effects of chloride or anion channel inhibitors on the initial rate of chloride uptake to the cytosol of zymosan-activated normal neutrophils. (A) Effects of chloride or anion channel inhibitors on the initial rate of chloride uptake to the cytosol of zymosan-activated neutrophils by colorimetric assay. Neutrophils, depleted of chloride by incubation in the NaGlu R for 1 h at 4°C, were allowed to phagocytose opsonized zymosan particles for 15 min at 37°C. The cells were then incubated with the stated concentrations of channel inhibitors in 135 mM NaCl R for 2, 5, and 10 min. Samples in triplicate were collected at each time-point and assayed for the total chloride content with a colorimetric assay as described in Materials and Methods. The initial rate of chloride uptake for each condition was calculated and compared with that of the DMSO control. The final concentration(s) for each inhibitor were GlyH-101 (CFTR inh , 12.5 μM, 25 μM, and 50 μM), CFTR inh 172 (CFTR inh , 20 μM), NA (100 μM), CHC (10 mM), DIDS (100 μM), and SITS (100 μM). *, P
Figure Legend Snippet: Effects of chloride or anion channel inhibitors on the initial rate of chloride uptake to the cytosol of zymosan-activated normal neutrophils. (A) Effects of chloride or anion channel inhibitors on the initial rate of chloride uptake to the cytosol of zymosan-activated neutrophils by colorimetric assay. Neutrophils, depleted of chloride by incubation in the NaGlu R for 1 h at 4°C, were allowed to phagocytose opsonized zymosan particles for 15 min at 37°C. The cells were then incubated with the stated concentrations of channel inhibitors in 135 mM NaCl R for 2, 5, and 10 min. Samples in triplicate were collected at each time-point and assayed for the total chloride content with a colorimetric assay as described in Materials and Methods. The initial rate of chloride uptake for each condition was calculated and compared with that of the DMSO control. The final concentration(s) for each inhibitor were GlyH-101 (CFTR inh , 12.5 μM, 25 μM, and 50 μM), CFTR inh 172 (CFTR inh , 20 μM), NA (100 μM), CHC (10 mM), DIDS (100 μM), and SITS (100 μM). *, P

Techniques Used: Colorimetric Assay, Incubation, Concentration Assay

33) Product Images from "Inhibition of sperm capacitation and fertilizing capacity by adjudin is mediated by chloride and its channels in humans"

Article Title: Inhibition of sperm capacitation and fertilizing capacity by adjudin is mediated by chloride and its channels in humans

Journal: Human Reproduction (Oxford, England)

doi: 10.1093/humrep/des384

Inhibition of human sperm capacitation by different blockers of Cl − channels. Sperm were preincubated with or without different blockers of Cl − channels, CFTRinh-172 (1 μM), adjudin (10 μM), DPC (10 μM), DIDs (10 μM) or/and bicuculline (Bic, 10 µM) for 5 h in the complete HTF medium under capacitating conditions. The results of capacitation (‘B’ + ‘AR’ patterns) are expressed as the mean ± SEM ( n = 5). a compared with control, P
Figure Legend Snippet: Inhibition of human sperm capacitation by different blockers of Cl − channels. Sperm were preincubated with or without different blockers of Cl − channels, CFTRinh-172 (1 μM), adjudin (10 μM), DPC (10 μM), DIDs (10 μM) or/and bicuculline (Bic, 10 µM) for 5 h in the complete HTF medium under capacitating conditions. The results of capacitation (‘B’ + ‘AR’ patterns) are expressed as the mean ± SEM ( n = 5). a compared with control, P

Techniques Used: Inhibition

34) Product Images from "Loss of MYO5B expression deregulates late endosome size which hinders mitotic spindle orientation"

Article Title: Loss of MYO5B expression deregulates late endosome size which hinders mitotic spindle orientation

Journal: PLoS Biology

doi: 10.1371/journal.pbio.3000531

vATPase and chloride channel activities mediate the late endosomal phenotype in Caco2 MYO5B−/− cells. (A–B) The numbers of vacuoles in Caco2 MYO5B –/– cells were reduced after treatments with vATPase inhibitor BafA1 and chloride channel inhibitors NPPB, DIDS, and furosemide for 24 h (A) and other appropriate times (B). Vacuoles reappeared after washout of the chemicals and culture in normal cell culture medium (B, arrow). Values for each data point can be found in S6 Data . (C–D) Caco2 MYO5B −/− cells revealed a shift in the abundance of the chloride channel ANO6 from the cell periphery to LAMP1-positive compartments (C, thick arrows) and vacuoles (D, thin arrows) compared with Caco2 WT cells. N = 3 independent experiments. Scale bars: 10 μm. ANO, anoctamin; Baf, bafilomycin; DIDS, 4,4'-Diisothiocyano-2,2'-stilbenedisulfonic acid; LAMP, late endosome–associated membrane protein; NPPB, 5-nitro-2-(3-phenylpropyl-amino) benzoic acid; vATPase, vacuolar H + -adenosine triphosphatase; WT, wild type.
Figure Legend Snippet: vATPase and chloride channel activities mediate the late endosomal phenotype in Caco2 MYO5B−/− cells. (A–B) The numbers of vacuoles in Caco2 MYO5B –/– cells were reduced after treatments with vATPase inhibitor BafA1 and chloride channel inhibitors NPPB, DIDS, and furosemide for 24 h (A) and other appropriate times (B). Vacuoles reappeared after washout of the chemicals and culture in normal cell culture medium (B, arrow). Values for each data point can be found in S6 Data . (C–D) Caco2 MYO5B −/− cells revealed a shift in the abundance of the chloride channel ANO6 from the cell periphery to LAMP1-positive compartments (C, thick arrows) and vacuoles (D, thin arrows) compared with Caco2 WT cells. N = 3 independent experiments. Scale bars: 10 μm. ANO, anoctamin; Baf, bafilomycin; DIDS, 4,4'-Diisothiocyano-2,2'-stilbenedisulfonic acid; LAMP, late endosome–associated membrane protein; NPPB, 5-nitro-2-(3-phenylpropyl-amino) benzoic acid; vATPase, vacuolar H + -adenosine triphosphatase; WT, wild type.

Techniques Used: Cell Culture

35) Product Images from "Exploring the role of stromal osmoregulation in cancer and disease using executable modelling"

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling

Journal: Nature Communications

doi: 10.1038/s41467-018-05414-y

Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a ) are deregulated in the model in the same manner as in the gene array. Experimental phenotype represents behaviour reported previously 42 , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P
Figure Legend Snippet: Application of the qualitative network to murine embryonic fibroblasts. Expansion of the QN to p53 −/− , Kras G12D/+ (HET) and p53 −/− , Kras G12D/G12D (HOM) MEFs. a Transport proteins deregulated within HOM MEFs when compared to HET MEFs. Proteins are upregulated (red arrows), downregulated (blue arrows), or unchanged (grey). b Phenotype change for MEFS when HOM MEFs are compared to HET MEFs. The model output represents the physiological behaviour predicted by the model when proteins from ( a ) are deregulated in the model in the same manner as in the gene array. Experimental phenotype represents behaviour reported previously 42 , where it is known. The model predicts that attachment will be significantly different between the two cell types, but viability and cell size will remain unchanged. c Effects of application of channel inhibitors to HOM MEFs in vitro showing application of 10 µM DIDs for 72 h, resulting in decreased cellular attachment (left), and application of 10 nM EIPA ± 10 µM DIDs (right). Data are technical replicates. Dotted lines represent calculated Bliss independence values. d Effects of application of 10 µM DIDs ± 10 nM EIPA on viability in HOM MEFs. e Effects of application of 10 µM DIDs ± 10 nM AHCL on attachment in HOM MEFs. f Effects of application of 10 µM DIDs ± 10 nM AHCL on viability in HOM MEFs.* P

Techniques Used: In Vitro, Cell Attachment Assay

36) Product Images from "Mechanisms of U46619-induced contraction of rat pulmonary arteries in the presence and absence of the endothelium"

Article Title: Mechanisms of U46619-induced contraction of rat pulmonary arteries in the presence and absence of the endothelium

Journal: British Journal of Pharmacology

doi: 10.1111/j.1476-5381.2008.00084.x

The effect of the potassium channel blockers PNU37883 (KATP ), ChTx (BKCa and IKCa ), TRAM 34 (IKCa ) and 4-AP (KV ) on basal tone and on the concentration–response curve to U46619 in E+ rings in the absence and presence of the VOCC blockers nifedipine and mibefradil, the chloride channel blockers NFA and DIDS or the putative IP3 receptor antagonist 2-APB
Figure Legend Snippet: The effect of the potassium channel blockers PNU37883 (KATP ), ChTx (BKCa and IKCa ), TRAM 34 (IKCa ) and 4-AP (KV ) on basal tone and on the concentration–response curve to U46619 in E+ rings in the absence and presence of the VOCC blockers nifedipine and mibefradil, the chloride channel blockers NFA and DIDS or the putative IP3 receptor antagonist 2-APB

Techniques Used: Concentration Assay

(A) The effect of VOCC and chloride channel blockade on KCl-induced constriction and (B) relaxation of U46619-induced tone by the chloride channel blockers DIDS and NFA compared with the BK Ca activator NS1619. (A) E+, the effect of nifedipine (1 µmol·L
Figure Legend Snippet: (A) The effect of VOCC and chloride channel blockade on KCl-induced constriction and (B) relaxation of U46619-induced tone by the chloride channel blockers DIDS and NFA compared with the BK Ca activator NS1619. (A) E+, the effect of nifedipine (1 µmol·L

Techniques Used:

The contractile response to the SERCA inhibitor CPA (10 µmol·L −1 ) alone in (E+) rat pulmonary arteries and in the presence of NPPB (30 µmol·L −1 ), NFA (30 µmol·L −1 ), DIDS (100 µmol·L
Figure Legend Snippet: The contractile response to the SERCA inhibitor CPA (10 µmol·L −1 ) alone in (E+) rat pulmonary arteries and in the presence of NPPB (30 µmol·L −1 ), NFA (30 µmol·L −1 ), DIDS (100 µmol·L

Techniques Used:

Effect of the chloride channel blockers NPPB, NFA and DIDS on the concentration–response curve for U46619-induced contraction of (E+) rat pulmonary arteries. (A) Response to U46619 in the absence and presence of NPPB at 10 µmol·L
Figure Legend Snippet: Effect of the chloride channel blockers NPPB, NFA and DIDS on the concentration–response curve for U46619-induced contraction of (E+) rat pulmonary arteries. (A) Response to U46619 in the absence and presence of NPPB at 10 µmol·L

Techniques Used: Concentration Assay

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MTT Assay:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Flow Cytometry:

Article Title: The Interaction between Trolox and 4,4’-diisothiocyanatostilbene-2,2’-disulfonic Acid on Hypoxic Pulmonary Vasoconstriction in the Isolated Rabbit Lung
Article Snippet: Subsequently, the lungs were randomly divided into four groups, namely control (HOX, n=5), DIDS (200 μM, Sigma, n=5) treated, Trolox (20 μM, Sigma, n=5) treated, and Trolox+DIDS treated group (n=5). .. PVR was calculated from pulmonary artery pressure, left atrial pressure, and perfusate flow rate by using Ohm equation.

esiRNA:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: 1000 Murine primary pmFRCs were reversely transfected with 100 ng esiRNA (Sigma) and seeded in triplicates into a 96-well plate. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Fluorescence:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: After 6 h of incubation under normal growth conditions, fluorescence intensities were measured at 560EX nm/590EM nm with a microplate reader (Tecan). .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons
Article Snippet: Fluorescence shorter than 490 nm was reflected by a long pass filter (NFT 490), and then filtered by a short pass filter with cutoff at 685 nm (KP685). .. After taking DIDS and DIC images, 10 μg/ml of propidium iodide (PI, Sigma, Ex/Em: 514/590 nm) was added, and images from the same view were taken 5 to 8 mins afterwards (excited by laser line 514 nm, and filtered by a long pass filter (LP560).

Synthesized:

Article Title: Molecular determinants of differential pore blocking of kidney CLC-K chloride channels
Article Snippet: .. DIDS was purchased from Sigma, and racemate 3-phenyl-CPP was synthesized in-house ( ). .. We thank T. Jentsch and R. Estévez for providing the CLC-Ka, CLC-Kb and barttin cDNA constructs; F. Loiodice, P. Tortorella and G. Fracchiolla for kindly providing 3-phenyl-CPP; E. Gaggero for the construction of the voltage-clamp amplifier and G. Gaggero for technical support in the construction of the voltage-clamp set-up.

Patch Clamp:

Article Title: Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes
Article Snippet: The K+ -free Tyrode's solution with the following composition [(in mM) 140 NaCl, 4 CsCl, 2 CaCl2 , 1 MgCl2 , 5 HEPES, 5 MES, 10 glucose and 10 sucrose at pH 7.4 (titrated with NaOH)] was superfused during all whole-cell patch clamp recordings. .. DIDS and Niflumic acid were purchased from Sigma (St. Louis, MO).

Generated:

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons
Article Snippet: After taking DIDS and DIC images, 10 μg/ml of propidium iodide (PI, Sigma, Ex/Em: 514/590 nm) was added, and images from the same view were taken 5 to 8 mins afterwards (excited by laser line 514 nm, and filtered by a long pass filter (LP560). .. Composite pictures were generated using Image J from DIC (grey), DIDS (blue), and PI (red).

Transferring:

Article Title: Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes
Article Snippet: For NMDG-Cl pipette solution, CsCl was totally replaced with equimolar NMDG-Cl. .. DIDS and Niflumic acid were purchased from Sigma (St. Louis, MO).

Transfection:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: 1000 Murine primary pmFRCs were reversely transfected with 100 ng esiRNA (Sigma) and seeded in triplicates into a 96-well plate. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Microscopy:

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons
Article Snippet: DIDS fluorescence (Ex/Em: 342/418 nm in water, with an emission Red-shift to > 450 nM when bound to protein ( )) was visualized by a Zeiss LSM510 META confocal laser scanning 2-photon microscope using a 40 X C-Apochromat (NA 1.2) water immersion objective. .. After taking DIDS and DIC images, 10 μg/ml of propidium iodide (PI, Sigma, Ex/Em: 514/590 nm) was added, and images from the same view were taken 5 to 8 mins afterwards (excited by laser line 514 nm, and filtered by a long pass filter (LP560).

Activation Assay:

Article Title: Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes
Article Snippet: The activation of ICl,pH was not different between these two conditions. .. DIDS and Niflumic acid were purchased from Sigma (St. Louis, MO).

Produced:

Article Title: Giardia secretome highlights secreted tenascins as a key component of pathogenesis
Article Snippet: Chemicals and Inhibitors Forskolin (10 μM), UTP (100 μM), Amiloride (10 μM), and DIDS (100 μM) were obtained from Sigma Aldrich, and GlyH-101 (50 μM) was obtained from Merck Chemicals. .. Final concentrations of drugs are as indicated in the text or figures and where produced by adding the appropriate volume of stock concentration to 5 ml of either the basolateral or apical bathing solution.

Concentration Assay:

Article Title: Molecular determinants of differential pore blocking of kidney CLC-K chloride channels
Article Snippet: For display, the I ( c )/ I (0) values at a given concentration were averaged and plotted versus concentration ( ). .. DIDS was purchased from Sigma, and racemate 3-phenyl-CPP was synthesized in-house ( ).

Article Title: PI3K and PKC contribute to membrane depolarization mediated by ?2-adrenoceptors in the canine isolated mesenteric vein
Article Snippet: Drugs NFA, NPPB, wortmannin, LY 294002, DIDS, nicardipine, norepinephrine hydrochloride, tetrodotoxin, phentolamine, xestospongin C, gadolinium chloride, PMA (Sigma-Aldrich), 2 APB (Tocris), PKCζPS and PKCζI (Calbiochem), chelerythrine, calphostin C (Biomol), general and phospho-specific anti-Akt antibodies (Cell Signaling Technology, Inc.), phosphatidylinositol (PI, Avanti). .. The final concentration of DMSO was less than 0.1 % DMSO.

Article Title: Salt secretion is linked to acid-base regulation of ionocytes in seawater-acclimated medaka: new insights into the salt-secreting mechanism
Article Snippet: Addition of DIDS, AZ, EIPA, and Bumetanide 4, 4′-diisothiocyanatostilbene-2, 2′-disulphonic acid (DIDS; 400 μM), acetazolamide (AZ; 200 μM), 5-(N-ethyl-N-isopropyl)-amiloride (EIPA; 400 μM), and bumetanide (Bumex; 400 μM) were obtained from Sigma-Aldrich, and were used to respectively suppress AE, CA, NHE, and NKCC in this study. .. The final concentration of DMSO or ethanol in the working solutions (including the control group) was 0.1%.

Article Title: Giardia secretome highlights secreted tenascins as a key component of pathogenesis
Article Snippet: Chemicals and Inhibitors Forskolin (10 μM), UTP (100 μM), Amiloride (10 μM), and DIDS (100 μM) were obtained from Sigma Aldrich, and GlyH-101 (50 μM) was obtained from Merck Chemicals. .. Final concentrations of drugs are as indicated in the text or figures and where produced by adding the appropriate volume of stock concentration to 5 ml of either the basolateral or apical bathing solution.

Article Title: Sat1 is dispensable for active oxalate secretion in mouse duodenum
Article Snippet: The apparent permeability coefficients ( P app ) for oxalate and mannitol were calculated using the following equation: J = P app × A × [S] where J is the flux, A is the cross-sectional tissue area of the Ussing chamber, and [S] is the substrate concentration of [14 C]oxalate or [3 H]mannitol. .. For experiments evaluating sensitivity to basolateral DIDS, 1 mM DIDS (Sigma-Aldrich, St. Louis, MO) was dissolved in oxygenated Ringer solution.

Incubation:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons
Article Snippet: Prior to experimentation, cells were rinsed once in serum-free DMEM and then incubated for 2-hrs with 0.2% DMSO or DIDS in DMEM or IS, and then rinsed once with DMEM or IS. .. After taking DIDS and DIC images, 10 μg/ml of propidium iodide (PI, Sigma, Ex/Em: 514/590 nm) was added, and images from the same view were taken 5 to 8 mins afterwards (excited by laser line 514 nm, and filtered by a long pass filter (LP560).

other:

Article Title: The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility
Article Snippet: Cl− channel modulators RIAA (Sigma-Aldrich), NPPB (Tocris Bioscience), and DIDS (Sigma-Aldrich) were used at the highest nontoxic concentrations: 50, 25, and 50 μm for HEK 293 cells, and 100, 50, 100 μm for MCC13 cells, respectively.

Article Title: The cellular chloride channels CLIC1 and CLIC4 contribute to virus-mediated cell motility
Article Snippet: RIAA (Sigma-Aldrich), NPPB (Tocris Bioscience), and DIDS (Sigma-Aldrich) were used at the highest nontoxic concentrations: 50, 25, and 50 μ m for HEK 293 cells, and 100, 50, 100 μ m for MCC13 cells, respectively.

Activity Assay:

Article Title: Sat1 is dispensable for active oxalate secretion in mouse duodenum
Article Snippet: Secretory fluxes of oxalate and mannitol were measured by adding 2 μM [14 C]oxalate (specific activity 117 mCi/mmol, Amersham Biosciences, Piscataway, NJ) and 0.08 μM [3 H]mannitol (specific activity 20 Ci/mmol, MP Biomedicals, Santa Ana, CA) to the serosal bath, and unlabeled 2 μM oxalate and 0.08 μM mannitol in the opposite bath. .. For experiments evaluating sensitivity to basolateral DIDS, 1 mM DIDS (Sigma-Aldrich, St. Louis, MO) was dissolved in oxygenated Ringer solution.

Cell Culture:

Article Title: Acidic pH-activated Cl- Current and Intracellular Ca2+ Response in Human Keratinocytes
Article Snippet: DIDS and Niflumic acid were purchased from Sigma (St. Louis, MO). .. Cell culture media, antibiotics and fetal bovine serum (FBS) were purchased from Gibco.

Article Title: SCRAMBLING OF PHOSPHOLIPIDS ACTIVATES RED CELL MEMBRANE CHOLESTEROL
Article Snippet: Human foreskin fibroblasts were cultured as described ( ). .. Ionomycin, TNBS, DIDS, CO ( Streptomyces sp ) and MBCD were all from Sigma.

Staining:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan). .. 5000 Murine primary pmFRCs were reversely transfected with 160 ng esiRNA (Sigma) and seeded in quadriplicate into a 96-well plate.

Permeability:

Article Title: Sat1 is dispensable for active oxalate secretion in mouse duodenum
Article Snippet: The apparent permeability coefficients ( P app ) for oxalate and mannitol were calculated using the following equation: J = P app × A × [S] where J is the flux, A is the cross-sectional tissue area of the Ussing chamber, and [S] is the substrate concentration of [14 C]oxalate or [3 H]mannitol. .. For experiments evaluating sensitivity to basolateral DIDS, 1 mM DIDS (Sigma-Aldrich, St. Louis, MO) was dissolved in oxygenated Ringer solution.

Viability Assay:

Article Title: Exploring the role of stromal osmoregulation in cancer and disease using executable modelling
Article Snippet: Twenty-four hours later the medium was exchanged and 72 h later cell viability was measured by the addition of CellTiter-Blue (Promega) viability assay reagent. .. Cells were then treated with indicated concentrations of DIDS, EIPA, AHCL or a combination of these and cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma) assay after 72 h. Cells were incubated with 0.5 mg/ml MTT solution for 3 h at 37 °C before stain was resuspended in DMSO and absorbance at 570 nm determined by microplate reader (Tecan).

Imaging:

Article Title: An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons
Article Snippet: Paragraph title: 4.4. DIDS permeation imaging ... After taking DIDS and DIC images, 10 μg/ml of propidium iodide (PI, Sigma, Ex/Em: 514/590 nm) was added, and images from the same view were taken 5 to 8 mins afterwards (excited by laser line 514 nm, and filtered by a long pass filter (LP560).

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    Millipore chemicals dids
    Chemicals Dids, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chemicals dids - by Bioz Stars, 2020-04
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