Figure 4. An ω-3 enriched diet increases TRPV4-mediated vasodilation in WT mice.

(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 Figure S4.