Figure 2: Wnt5a promotes AQP2 trafficking through the activation of calcium signalling.

(a) The subcellular localization of total AQP2 and phosphorylated AQP2 at S269. The mpkCCD cells were treated with Wnt5a (500 ng ml⁻¹) for 1 h, and the subcellular localization of AQP2 was then analysed by immunofluorescence using confocal microscopy. The larger panels display confocal sections of the apical regions of the cells. Z-stack confocal images are shown at the top of each panel. Representative confocal images of three independent experiments are shown. Scale bars, 10 μm. (b) The effects of W7, CyA and KN93 on Wnt5a-induced AQP2 trafficking. Wnt5a (500 ng ml⁻¹) was added in the presence or absence of W7 (100 μM), CyA (10 μM), or KN93 (10 μM) for 1 h. The mpkCCD cells were pretreated with each inhibitor for 45 min before Wnt5a stimulation. Immunofluorescence staining of AQP2 was analysed as in a. Scale bars, 10 μm. Fluorescence intensities of apical AQP2 were quantified, and the results are presented in the bar graphs. Error bars are mean values±s.d. from three experiments. Tukey, **P<0.01.