Fig 1.
Knockdown of TRPV4 with siRNA in HTMC.
(A) The knockdown of TRPV4 mRNA was confirmed by qPCR. Data are mean ± SE (n = 3). (B) Representative pseudo-color images of intracellular Ca2+ changes in control or TRPV4 siRNA-treated HTMC before and after stimulation with TRPV4 agonist GSK1016790A. Scale bar, 100 μm. (C, D) Quantification of GSK1016790A-induced intracellular Ca2+ elevation (change in Fluo-8 ratio) in control or TRPV4 siRNA-treated HTMC. Black bar represents GSK1016790A treatment. Data are expressed as the mean ± SE (n = 5).
Fig 2.
The effect of TRPV4 knockdown on intracellular Ca2+ changes induced by extension stimuli in HTMC.
(A) Representative pseudo-color images of intracellular Ca2+ changes in control or TRPV4 siRNA-treated HTMC before and after mechanical extension stimuli. Scale bar, 100 μm. (B) Quantification of mechanical extension stimuli-induced intracellular Ca2+ elevation (change in Fluo-8 ratio) in control or TRPV4 siRNA-treated HTMC. Data are expressed as the mean ± SE (n = 5).
Fig 3.
Effect of TRPV4 knockdown on enhanced arachidonic acid and PGE2 release by stretch in HTMC.
The amount of arachidonic acid (A) and PGE2 (B) in cell supernatant 10 min after mechanical stretch stimulation in control or TRPV4 siRNA-treated HTMC. Data are expressed as mean ± SE (n = 3).
Fig 4.
GSK1016790A promotes the release of PGD2 in addition to arachidonic acid and PGE2 from HTMC.
The amount of arachidonic acid (A), PGE2 (B) and PGD2 (C) in the culture supernatant of HTMC at each time after GSK1016790A treatment. Data are expressed as the mean ± SE (n = 3).
Fig 5.
Effects of GSK1016790A on contraction of HTMC.
Change rate in area of the collagen gels in presence of DMSO, 10 or 100 nM GSK1016790A were measured. Data are expressed as the mean ± SE (n = 4).
Fig 6.
TRPV4 regulates intraocular pressure in mice.
(A) Intraocular pressure (IOP) in nine to twelve-week-old TRPV4+/+ and TRPV4−/− mice. IOP was measured using the microneedle method. Data are expressed as the mean ± SE (n = 22–24). (B) TRPV4 agonist GSK1016790A lowers IOP in C57BL/6 Wild type mice. Mice were treated with vehicle (0.5% DMSO) or 0.25 mg/kg GSK1016790A. IOP was measured using the microneedle method 1.5 h after intraperitoneal injection. Data are expressed as the mean ± SE (n = 22).