FIGURE 7.

TRPV4 activation increased actin polymerization and cell adhesion. A, fluorescent photomicrograph of phalloidin, a marker for F-actin. In the presence of NGF, treatment with 10 μm 4α-PDD or 300 nm 5,6-EET induced morphological change and increased phalloidin fluorescence intensities. Phalloidin was conjugated with Alexa Fluor 488. Nuclei were stained with Hoechst 33342. Scale bar, 10 μm. NGF and TRPV4 activators were applied to cells for 24 h before fixation for staining. B, after shaking, numbers of attached cells were markedly greater after treating cells with 4α-PDD or 5,6-EET. Cell numbers were determined by staining nuclei with Hoechst 33342. C, quantification of phalloidin fluorescence intensities. 4α-PDD and 5,6-EET increased the phalloidin fluorescence intensities of cells treated with or without NGF. D, summary of the effect of treatment with TRPV4 agonists (4α-PDD or 5,6-EET) on numbers of attached cells. Note that TRPV4 activators significantly increased the number of cells attached to plates after shaking.