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Fig. 3. TRPV4 is necessary for regulatory volume decrease. Relative volume changes of HaCaT cells and CHO cells exposed to hypotonic or isotonic medium at t=0 seconds. (A) Reduction of extracellular osmolarity led to a rapid increase in cell volume of untreated HaCaT cells (hypo, filled circles, n=18) followed by RVD. Replacement of the medium alone did not affect cell volume (iso, open circles, n=12). Treatment with 100 µM Gd³⁺ (filled triangles, n=8) or Ca²⁺-free solution (open squares, n=8) abolished the RVD response and cell volume stayed elevated. (B) Untransfected CHO cells (nt, open circles, n=22) showed no RVD, whereas TRPV4-transfected CHO cells (TRPV4, filled circles, n=25) were able to undergo RVD. Under Ca²⁺-free conditions (open squares, n=8), no RVD occurred after the initial swelling in transfected cells. Gd³⁺-pretreated transfected cells also showed no RVD in comparison with untreated cells (filled triangles, n=14). Cell volume was estimated from the central cross-sectional area and normalized to control (t $≤$ 0 seconds) values. Data are means±s.e.m.

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