rabbit polyclonal anti-human trpv4  (WuXi AppTec)

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) Top: representative current-voltage relationships determined by whole-cell patch-clamp recordings of control and EPA (100 μM)-treated aortic endothelial cells challenged with GSK1016790A (GSK101, TRPV4 agonist, 0.1 μM) and GSK101 (0.1 μM) + HC067047 (HC, TRPV4 antagonist; 5 μM). Bckgrd. indicates background currents. Bottom: representative time course of whole-cell patch-clamp recordings (+80 mV) of control and EPA-treated aortic endothelial cells challenged with GSK101 and inhibited with HC. (B) Boxplots show the mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of TRPV4 currents ((I GSK101 – I HC ) pA/pF) obtained by whole-cell patch-clamp recordings (+80 mV) of control and EPA-treated endothelial cells from skin, retina, lung, brain, and aorta. Two-way ANOVA and Sidak-Holm multiple comparisons test. (C) Left: representative current-voltage relationships determined by whole-cell patch-clamp recordings of control and EPA (100 μM)-treated aortic endothelial cells challenged with GSK101 (from 1 to 2,000 nM). Currents evoked by GSK101 submaximal concentrations (gray and red traces) were normalized by corresponding currents elicited by saturating GSK101 (2,000 nM; black traces) per cell. Right: normalized (I/I max ) GSK101 dose-response profiles of control and EPA (100 μM)-treated aortic endothelial cells. A Hill function was fitted to the data. The shadows encompassing the curves indicate the 95% confidence bands for the fit. Circles are mean ± SD. n = 36 for control and n = 36 for EPA (100 μM)-treated aortic endothelial cells. (D) Top: representative western blots (anti-TRPV4) of the membrane fractions of control and EPA (100 μM)-treated human endothelial cells from skin, retina, lung, brain, and aorta. Bottom: mean/scatter-dot plot showing relative intensities of TRPV4 bands, calculated from total protein detection of chemically labeled proteins (Stain-Free System Bio-Rad), from the membrane fractions of control and EPA (100 μM)-treated endothelial cells. Lines are mean ± SD. Two-way ANOVA. Asterisks indicate values significantly different from control (***p < 0.001 and **p < 0.01) and n.s. indicates values not significantly different from the control. n is indicated in each panel. See also and .

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Patch Clamp, Western Blot, Labeling, Staining

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) The C. elegans fatty acid desaturase FAT-1 enzyme introduces a double bond in ω -6 arachidonic acid to synthesize ω -3 EPA in worms and transgenic fat-1 mice, but not in WT mice. Mice and C. elegans cartoons were created with BioRender.com . (B) Gardner-Altman estimation plot showing the mean difference in EPA-membrane content of whole mesenteric arteries of WT and fat -1 mice, as determined by LC-MS. The raw data are plotted on the left axis. The mean difference, on the right, is depicted as a dot; the 95% confidence interval is indicated by the ends of the vertical error bars. Mann-Whitney rank test for two independent groups. (C) Representative current-voltage relationships determined by whole-cell patch-clamp recordings of WT and fat -1 cultured isolated mesenteric endothelial cells challenged with GSK101 (0.1 μM) and GSK101 (0.1 μM) + HC (10 μM). Bckgrd. indicates background currents. (D) Bar graph displaying TRPV4 currents ((I GSK101 – I HC ) pA/pF) obtained by whole-cell patch-clamp recordings (+80 mV) of cultured isolated mesenteric endothelial cells of WT and fat -1 mice. Bars are mean ± SEM. Two-tailed unpaired t test. Asterisks indicate values significantly different from WT (**p < 0.01 and *p < 0.05). n is indicated in each panel.

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Transgenic Assay, Liquid Chromatography with Mass Spectroscopy, MANN-WHITNEY, Patch Clamp, Cell Culture, Isolation, Two Tailed Test

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) Representative time course of GSK101 (5 nM)-induced vasodilation of pressurized (80 mmHg) mesenteric arteries from WT and fat-1 mice. Inset: micrograph of a representative cannulated mesenteric artery. (B) Percentage of GSK101 (5 nM)-induced vasodilation of mesenteric arteries (endothelium-intact or -denuded) from WT and fat-1 mice. Bars are mean ± SEM. Two-way ANOVA and Tukey multiple comparisons test. (C) Representative western blot (anti-TRPV4) of the membrane fractions of WT and fat-1 mice mesenteric arteries. (D) Mean/scatter-dot plot showing relative intensities of TRPV4 bands, calculated from total protein detection of chemically labeled proteins (Stain-Free System Bio-Rad), from the membrane fractions of mesenteric arteries from WT and fat-1 mice. Lines are mean ± SD. Two-tailed unpaired t test. Asterisks indicate values significantly different from WT (**p < 0.01) and n.s. indicates values not significantly different from the WT. n is indicated in each panel. See also .

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Western Blot, Labeling, Staining, Two Tailed Test

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) Gardner-Altman estimation plot showing the mean difference in EPA-membrane content of whole mesenteric arteries of WT mice fed with standard or ω -3 fatty acid-enriched diets, as determined by LC-MS. The raw data are plotted on the left axis. The mean difference, on the right, is depicted as a dot; the 95% confidence interval is indicated by the ends of the vertical error bars. Mann-Whitney rank test for two independent groups. Mouse cartoon was created with BioRender.com . (B) Representative time course of GSK101 (5 nM)-induced vasodilation of pressurized (80 mmHg) mesenteric arteries from WT mice fed with standard or ω -3 fatty acid-enriched diets. (C) Percentage of GSK101 (5 nM)-induced vasodilation of mesenteric arteries from WT mice fed with standard or ω -3 fatty acid-enriched diets. Bars are mean ± SEM. Two-tailed unpaired t test. (D) Boxplots show the mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of the percentage of maximal GSK101 (5 nM)-induced vasodilation remaining at the time GSK101 was removed from the mesenteric arteries of WT mice fed with standard or ω -3 fatty acid-enriched diets. Two-tailed unpaired t test. (E) Top: representative western blot (TRPV4) from membrane fractions of the mesenteric arteries from WT mice fed with standard or ω -3 fatty acid-enriched diets. Bottom: mean/scatter-dot plot showing relative intensities of TRPV4 bands, calculated from total protein detection of chemically labeled proteins (Stain-Free System Bio-Rad), from the membrane fractions of mesenteric arteries from WT mice fed with standard or ω -3 fatty acid-enriched diets. Lines are mean ± SD. Two-tailed unpaired t test. (F) Representative current-voltage relationships determined by whole-cell patch-clamp recordings of cultured isolated mesenteric endothelial cells, from WT mice fed with standard or ω -3 fatty acid-enriched diets, challenged with GSK101 (0.1 μM) and GSK101 (0.1 μM) + HC (10 μM). (G) Bar graph displaying TRPV4 currents ((I GSK101 – I HC ) pA/pF) obtained by whole-cell patch-clamp recordings (+80 mV) of cultured isolated mesenteric endothelial cells of WT mice fed with standard or ω -3 fatty acid-enriched diets. Bars are mean ± SEM. Two-tailed unpaired t test. (H) Top: representative time course of sodium nitroprusside (SNP) (1 μM)-induced vasodilation of pressurized (80 mmHg) mesenteric arteries from WT mice fed with standard or ω -3 fatty acid-enriched diets. Bottom: percentage of SNP (1 μM)-induced vasodilation of WT mice fed with standard or ω -3 fatty acid-enriched diets. Bars are mean ± SEM. Two-tailed unpaired t test. Asterisks indicate values significantly different from standard diet (***p < 0.001, **p < 0.01, and *p < 0.05) and n.s. indicates values not significantly different from the standard diet. n is indicated in each panel. See also .

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Liquid Chromatography with Mass Spectroscopy, MANN-WHITNEY, Two Tailed Test, Western Blot, Labeling, Staining, Patch Clamp, Cell Culture, Isolation

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) Representative time course of whole-cell patch-clamp recordings (−60 mV) of control and EPA (100 μM)-treated human aortic endothelial cells challenged with GSK101 (1 μM). (B) Bar graph displaying TRPV4 currents (pA/pF) obtained by whole-cell patch-clamp recordings (−60 mV) of control and EPA (100 μM)-treated aortic endothelial cells. Bars are mean ± SEM. Mann-Whitney rank test for two independent groups. (C) Boxplots show mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of the time to reach half amplitude from the peak current (Imax) elicited by GSK101 (1 μM) in control and EPA (100 μM)-treated aortic endothelial cells. Two-tailed unpaired t test. (D) Micrographs of cultured isolated mesenteric endothelial cells from WT mice fed with standard or ω -3 fatty acid-enriched diets, loaded with Fluo-4 AM, and challenged with GSK101 (0.1 μM). The color bar indicates a relative change in fluorescence intensity. Experiments were performed in three independent cell preparations. t indicates the times at which representative micrographs were taken from the traces in (E). (E) Representative traces corresponding to normalized intensity changes (ΔF/F) of individual cells shown in (D). (F) Area under the curve (AUC) of the fluorescence response (ΔF/F), depicted as violin plots with the means shown as horizontal bars, of endothelial cells from WT mice fed with standard or ω -3 fatty acid-enriched diets challenged with GSK101. Two-tailed unpaired t test. (G) Micrographs of cultured isolated mesenteric endothelial cells of WT mice fed with standard or ω -3 fatty acid-enriched diets, loaded with Fluo-4 AM and challenged with a hypoosmotic buffer (HB: 240 mOsm). The color bar indicates a relative change in fluorescence intensity. Experiments were performed in two independent cell preparations. t indicates the times at which representative micrographs were taken from the traces in (H). (H) Representative traces corresponding to normalized intensity changes (ΔF/F) of individual cells shown in (G). (I) AUC of the fluorescence response (ΔF/F), depicted as violin plots with the means shown as horizontal bars, of endothelial cells from WT mice fed with standard or ω -3 fatty acid-enriched diets challenged with a hypoosmotic buffer. Two-tailed unpaired t test. Asterisks indicate values significantly different from control or standard diet (***p < 0.001 and **p < 0.01). n is indicated in each panel.

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Patch Clamp, MANN-WHITNEY, Two Tailed Test, Cell Culture, Isolation, Fluorescence

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: (A) Cartoon depicting the full-length rat TRPV4 channel. (B) Representative time course of whole-cell patch-clamp recordings (−60 mV) of control and EPA (100 μM)-treated HEK293 cells, expressing full-length rat TRPV4, challenged with GSK101 (1 μM). Traces were normalized for comparison. (C) Boxplots show mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of the time to reach half amplitude from the peak current (Imax) elicited by GSK101 (1 μM) in control and EPA (100 μM)-treated HEK293 cells expressing full-length rat TRPV4. Two-tailed unpaired with Welch’s correction. (D) Cartoon depicting the Δ186 rat TRPV4 channel construct. (E) Representative time course of whole-cell patch-clamp recordings (−60 mV) of control and EPA (100 μM)-treated HEK293 cells, expressing the Δ186 rat TRPV4 channel construct, challenged with GSK101 (1 μM). Traces were normalized for comparison. (F) Boxplots show mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of the time to reach half amplitude from the peak current (Imax) elicited by GSK101 (1 μM) in control and EPA (100 μM)-treated HEK293 cells expressing the Δ186 rat TRPV4 channel construct. Two-tailed unpaired t test. (G) Cartoon depicting the 5Ala (K121A, R122A, R124A, R125A, K126A) rat TRPV4 channel construct. (H) Representative time course of whole-cell patch-clamp recordings (−60 mV) in control and EPA (100 μM)-treated HEK293 cells expressing the 5Ala rat TRPV4 channel construct challenged with GSK101 (1 μM). Traces were normalized for comparison. (I) Boxplots show mean (gray circle), median (bisecting line), SD (whiskers), and SEM (box) of the time to reach half amplitude from the peak current (Imax) elicited by GSK101 (1 μM) of control and EPA (100 μM)-treated HEK293 cells expressing the 5Ala rat TRPV4 channel construct. Two-tailed unpaired t test. Asterisks indicate values significantly different from control (**p < 0.01) and n.s. indicates values not significantly different from the control. n is indicated in each panel. See also .

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Patch Clamp, Expressing, Two Tailed Test, Construct

Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-human TRPV4 , Abcepta , Cat# AP18990a, RRID: AB_2923215.

Techniques: Recombinant, Protease Inhibitor, Staining, Western Blot, Protein Extraction, Plasmid Preparation, Software, Imaging, Microscopy