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Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

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Structured Review

Bio-Techne corporation gap27
Cx43-formed hemichannels, but not gap junction channels, are associated with endothelial cell migration. a , Representative images of dye coupling assay (left) and the analysis of the number of coupled cells via gap junction communication (right) attained in the intact monolayer and in the migration front after scratching the monolayer. Dye coupling was assessed by measuring after 2 min the diffusion to neighboring cells (coupled cells) of the ethidium bromide microinjected into a single endothelial cell. The yellow diamond indicates the microinjected cell. b , Representative images of the ethidium uptake observed in primary cultures of mesenteric endothelial cells in control conditions and in the presence of the Cx blocking peptide 37,43 <t>Gap27</t> (200 µM) or the Cx43 hemichannel inhibitor TAT-Gap19 (Gap19, 300 µM) (left). Ethidium uptake was evaluated 15 min after scratching the monolayer and cells were incubated with the dye for 15 min, as shown in the time course of ethidium uptake observed in the intact monolayer and in the migration front (b, right top). Dot yellow lines depict the edge of the migration front. In addition, the analysis of the ethidium uptake rate achieved in the intact monolayer and in the migration front in control conditions and in the presence of 37,43 Gap27 or Gap19 is also shown (b, right bottom). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. Changes in ethidium-fluorescence signal are expressed in arbitrary units (a.u.). Numbers inside the bars indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Monolayer by two-way ANOVA. ††, P < 0.01 and †††, P < 0.001 vs. Migration front in control conditions (Control) by one-way ANOVA plus Bonferroni post hoc test. &, P < 0.001 vs. Migration front by paired Student’s t-test
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images

1) Product Images from "Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration"

Article Title: Opening of Cx43-formed hemichannels mediates the Ca 2+ signaling associated with endothelial cell migration

Journal: Biology Direct

doi: 10.1186/s13062-023-00408-3

Cx43-formed hemichannels, but not gap junction channels, are associated with endothelial cell migration. a , Representative images of dye coupling assay (left) and the analysis of the number of coupled cells via gap junction communication (right) attained in the intact monolayer and in the migration front after scratching the monolayer. Dye coupling was assessed by measuring after 2 min the diffusion to neighboring cells (coupled cells) of the ethidium bromide microinjected into a single endothelial cell. The yellow diamond indicates the microinjected cell. b , Representative images of the ethidium uptake observed in primary cultures of mesenteric endothelial cells in control conditions and in the presence of the Cx blocking peptide 37,43 Gap27 (200 µM) or the Cx43 hemichannel inhibitor TAT-Gap19 (Gap19, 300 µM) (left). Ethidium uptake was evaluated 15 min after scratching the monolayer and cells were incubated with the dye for 15 min, as shown in the time course of ethidium uptake observed in the intact monolayer and in the migration front (b, right top). Dot yellow lines depict the edge of the migration front. In addition, the analysis of the ethidium uptake rate achieved in the intact monolayer and in the migration front in control conditions and in the presence of 37,43 Gap27 or Gap19 is also shown (b, right bottom). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. Changes in ethidium-fluorescence signal are expressed in arbitrary units (a.u.). Numbers inside the bars indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Monolayer by two-way ANOVA. ††, P < 0.01 and †††, P < 0.001 vs. Migration front in control conditions (Control) by one-way ANOVA plus Bonferroni post hoc test. &, P < 0.001 vs. Migration front by paired Student’s t-test
Figure Legend Snippet: Cx43-formed hemichannels, but not gap junction channels, are associated with endothelial cell migration. a , Representative images of dye coupling assay (left) and the analysis of the number of coupled cells via gap junction communication (right) attained in the intact monolayer and in the migration front after scratching the monolayer. Dye coupling was assessed by measuring after 2 min the diffusion to neighboring cells (coupled cells) of the ethidium bromide microinjected into a single endothelial cell. The yellow diamond indicates the microinjected cell. b , Representative images of the ethidium uptake observed in primary cultures of mesenteric endothelial cells in control conditions and in the presence of the Cx blocking peptide 37,43 Gap27 (200 µM) or the Cx43 hemichannel inhibitor TAT-Gap19 (Gap19, 300 µM) (left). Ethidium uptake was evaluated 15 min after scratching the monolayer and cells were incubated with the dye for 15 min, as shown in the time course of ethidium uptake observed in the intact monolayer and in the migration front (b, right top). Dot yellow lines depict the edge of the migration front. In addition, the analysis of the ethidium uptake rate achieved in the intact monolayer and in the migration front in control conditions and in the presence of 37,43 Gap27 or Gap19 is also shown (b, right bottom). The rate of ethidium uptake was assessed by calculating the slope of the increase in fluorescence intensity along the time. Changes in ethidium-fluorescence signal are expressed in arbitrary units (a.u.). Numbers inside the bars indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Monolayer by two-way ANOVA. ††, P < 0.01 and †††, P < 0.001 vs. Migration front in control conditions (Control) by one-way ANOVA plus Bonferroni post hoc test. &, P < 0.001 vs. Migration front by paired Student’s t-test

Techniques Used: Migration, Diffusion-based Assay, Blocking Assay, Incubation, Fluorescence

Endothelial cell migration depends on a Cx43-formed channel-mediated increase in intracellular Ca 2+ concentration ([Ca 2+ ] i ). a , Representative images (left) and fluorescence intensity analysis (right) of the increase in [Ca 2+ ] i observed in endothelial cells of the migration front 15 min after scratching the monolayer in control conditions and in the presence of 50 µM 18-β-Glycyrrhetenic acid (ß-GA), a general Cx-formed channel blocker, or 200 µM 37,43 Gap27, a peptide designed to block channels formed by Cx37 or Cx43. Variations in the levels of [Ca 2+ ] i were assessed with the fluorescent Ca 2+ indicator Fluo-4. b , Representative images of the changes in [Ca 2+ ] i of endothelial cells (left) in which the differences in the subcellular distribution of the Ca 2+ signal attained in a cell of the migration front (Cell 2) and a cell within the monolayer (Cell 1) are highlighted in a 3D analysis (middle and right). c , Fluorescence intensity analysis of the Fluo-4 signal measured along the endothelial cells length (from back to front) in the migration front and the monolayer. d , Analysis of the changes in [Ca 2+ ] i levels attained in the rear and anterior edge of endothelial cells of the migration front in control conditions and after the treatment with ß-GA or 37,43 Gap27. Changes in Fluo-4 signal are expressed as the area under the curve (AUC). Note that cells were treated with the Cx blocking peptide 37,43 Gap27 for only 10 min to inhibit Cx hemichannels, without affecting gap junction channels. Dot red lines depict the edge of the migration front. Numbers inside the bars or in parentheses indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Control by one-way ANOVA plus Bonferroni post hoc test. †††, P < 0.001 vs. Anterior by paired Student’s t-test
Figure Legend Snippet: Endothelial cell migration depends on a Cx43-formed channel-mediated increase in intracellular Ca 2+ concentration ([Ca 2+ ] i ). a , Representative images (left) and fluorescence intensity analysis (right) of the increase in [Ca 2+ ] i observed in endothelial cells of the migration front 15 min after scratching the monolayer in control conditions and in the presence of 50 µM 18-β-Glycyrrhetenic acid (ß-GA), a general Cx-formed channel blocker, or 200 µM 37,43 Gap27, a peptide designed to block channels formed by Cx37 or Cx43. Variations in the levels of [Ca 2+ ] i were assessed with the fluorescent Ca 2+ indicator Fluo-4. b , Representative images of the changes in [Ca 2+ ] i of endothelial cells (left) in which the differences in the subcellular distribution of the Ca 2+ signal attained in a cell of the migration front (Cell 2) and a cell within the monolayer (Cell 1) are highlighted in a 3D analysis (middle and right). c , Fluorescence intensity analysis of the Fluo-4 signal measured along the endothelial cells length (from back to front) in the migration front and the monolayer. d , Analysis of the changes in [Ca 2+ ] i levels attained in the rear and anterior edge of endothelial cells of the migration front in control conditions and after the treatment with ß-GA or 37,43 Gap27. Changes in Fluo-4 signal are expressed as the area under the curve (AUC). Note that cells were treated with the Cx blocking peptide 37,43 Gap27 for only 10 min to inhibit Cx hemichannels, without affecting gap junction channels. Dot red lines depict the edge of the migration front. Numbers inside the bars or in parentheses indicate the n value. Values are means ± SEM. ***, P < 0.001 vs. Control by one-way ANOVA plus Bonferroni post hoc test. †††, P < 0.001 vs. Anterior by paired Student’s t-test

Techniques Used: Migration, Concentration Assay, Fluorescence, Blocking Assay


Structured Review

Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images


Structured Review

Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images


Structured Review

Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images


Structured Review

Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images


Structured Review

Bio-Techne corporation gap27
( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of <t>GAP27</t> (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images

1) Product Images from "A succinate/SUCNR1-brush cell defense program in the tracheal epithelium"

Article Title: A succinate/SUCNR1-brush cell defense program in the tracheal epithelium

Journal: Science Advances

doi: 10.1126/sciadv.adg8842

( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Figure Legend Snippet: ( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.

Techniques Used: Activity Assay, Transmission Assay

( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).
Figure Legend Snippet: ( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).

Techniques Used: Inhibition, MANN-WHITNEY

( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.
Figure Legend Snippet: ( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.

Techniques Used: MANN-WHITNEY


Structured Review

Bio-Techne corporation gap27
( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of <t>GAP27</t> (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

Images

1) Product Images from "A succinate/SUCNR1-brush cell defense program in the tracheal epithelium"

Article Title: A succinate/SUCNR1-brush cell defense program in the tracheal epithelium

Journal: Science Advances

doi: 10.1126/sciadv.adg8842

( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Figure Legend Snippet: ( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.

Techniques Used: Activity Assay, Transmission Assay

( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).
Figure Legend Snippet: ( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).

Techniques Used: Inhibition, MANN-WHITNEY

( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.
Figure Legend Snippet: ( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.

Techniques Used: MANN-WHITNEY


Structured Review

Bio-Techne corporation gap27
( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of <t>GAP27</t> (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

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1) Product Images from "A succinate/SUCNR1-brush cell defense program in the tracheal epithelium"

Article Title: A succinate/SUCNR1-brush cell defense program in the tracheal epithelium

Journal: Science Advances

doi: 10.1126/sciadv.adg8842

( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.
Figure Legend Snippet: ( A ) A model of succinate-evoked Ca 2+ wave propagation in the trachea that predicts sequential activity of cholinergic transmission and long-range gap junction communication (see explanation in text). TRPM5 + trigger cell, red; succinate-evoked Ca 2+ activity in epithelial cells, light green. ( B and C ) Example of the spatiotemporal properties of succinate-evoked Ca 2+ waves and the effects of GAP27 (130 μM) or a combination of GAP27 and 4-DAMP (1 μM). Borders of cells, light gray. Time course analyses of individual cells within white box (B) are depicted in fig. S6 (A and B). Arrows, TRPM5 + cells. ( D ) Group data of experiments as shown in (C). Friedman analysis of variance (ANOVA), Dunn’s. Data, median ± interquartile range (IQR). ( E to I ) Original examples and group data showing succinate-evoked Ca 2+ responses in TRPM5 + cells before and after treatment with GAP27 (130 μM), TAT-GAP19 (100 μM), or 4-DAMP (1 μM). Paired t test. Data, mean ± SD. ( J and K ) Δ F / F density peak values (mean ± SD) of succinate-evoked Ca 2+ waves after treatment with either apyrase mix (apyrase VI and VII, 5 U/ml each) or pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM). Succinate, 1 mM; adenosine triphosphate (ATP), 20 μM. ANOVA, posthoc: Tukey. Numbers in parentheses indicate independent experiments.

Techniques Used: Activity Assay, Transmission Assay

( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).
Figure Legend Snippet: ( A to F ) Maximum changes in particle transport speed (PTS) evoked by succinate in tracheae with brush cell–specific deletion of the acetylcholine (ACh)–synthesizing enzyme Chat ( Avil Cre Chat fl/fl ) (A) or global deletion of the M3 muscarinic receptor ( Chrm3 −/− ) (B), under general muscarinic receptor blockade with atropine (10 μM) (C), gap junction inhibition by GAP27 (130 μM, given 45 min before succinate) (D), SERCA inhibition by cyclopiazonic acid (CPA) (30 μM) (E), P2X4 and P2X7 inhibition by 5-BDBD (10 μM) and A438079 (20 μM) (E), general P2 receptor inhibition by pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) (100 μM) (F), and apyrase treatment (apyrase VI and VII, 5 U/ml each) (F). Control conditions were heterozygous littermates ( Avil Cre Chat fl/+ ) (A), homozygous wild-type animals from the same strain ( Chrm3 +/+ ) (C), and respective vehicle treatment [(B) and (D) to (F)]. Mann-Whitney test. Data, median ± interquartile range (IQR). Succinate, 1 mM; adenosine triphosphate (ATP), 100 μM, if not indicated otherwise (D). Numbers in parentheses indicate independent experiments (= tracheae).

Techniques Used: Inhibition, MANN-WHITNEY

( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.
Figure Legend Snippet: ( A , B , D , E , G , H , J , and K ) Representative recordings of transepithelial short-circuit current ( I SC ) in an opened mouse trachea mounted in an Ussing chamber and exposed to succinate (1 mM), ( C , F , I , and L ) group data showing maximum increases in I SC (Δ I SC ). The adenylate cyclase activator forskolin (10 μM, given apically) served as vitality control throughout. Vehicle control for experiments with the muscarinic M3 receptor blocker 4-DAMP (1 μM) [(A) to (C)], the SERCA-inhibitor cyclopiazonic acid (CPA) (30 μM) [(D) to (F)], and the gap junction inhibitor GAP27 (130 μM, given 45 min before succinate) [(G) to (I)] was DMSO [(A) to (C), 1 mM; (D) to (I), 500 μM]. Apyrase was given as a mix of apyrase VI and VII, 5 U/ml each [(J) to (L)]. All inhibitors were applied both apically and basolaterally. Data are from three independent experiments in (C), from two in (F) and (I), and from five in (L). Data, median ± interquartile range (IQR), number of tracheae is given in parentheses; Mann-Whitney test.

Techniques Used: MANN-WHITNEY


Structured Review

Bio-Techne corporation gap27
Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gap27/product/Bio-Techne corporation
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gap27 - by Bioz Stars, 2024-04
86/100 stars

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    Bio-Techne corporation gap27
    Gap27, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gap27/product/Bio-Techne corporation
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gap27 - by Bioz Stars, 2024-04
    86/100 stars
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