Changes of subcellular localization of caveolins after hypo-osmotic stress (related to Fig. 1 ). (A) Wild-type EpH4 cells stained with anti-Claudin-3 pAb and anti-Caveolin-1 mAb. The lower panels show the snapshots in the z-axis direction. Scale bar, 10 µm. (B) Wild-type EpH4 cells stably expressed cGFP-tagged Caveolin-2. The lower pictures show the snapshot in the z-axis direction. Scale bar, 5 µm. (C) Time-lapse imaging of EpH4 cells stably expressing Caveolin-2-GFP. Hypo-osmotic buffer (150 mOsm/liter) was added at time 0:00. Scale bar, 5 µm. (D) Fluorescence intensities normalized to the value at t = 0 min and shown as the mean line. N = 3 independent experiments. (E) Wild-type MDCK cells stained with anti-Caveolin-1 mAb. The lower panels show the snapshots in the z-axis direction. Scale bar, 10 µm. (F) Wild-type MDCK cells treated with hypo-osmotic buffer (150 mOsm/liter) for indicated time were fixed and stained with anti-Claudin-3 pAb and anti-Caveolin mAb. Scale bar, 10 µm. (G) Quantification of the ratio of fluorescence signal of caveolin-1 staining of the apical membrane to that of the basolateral membrane in wild-type MDCK cells treated with hypo-osmotic buffer (150 mOsm/liter) for indicated time. N = 4 independent experiments; error bar, SD; ns, not significant by one-way ANOVA with Tukey’s post-hoc test.