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. 2008 Apr 23;295(1):F290–F294. doi: 10.1152/ajprenal.00072.2008

Fig. 3.

Fig. 3.

VP stimulated AQP2 trafficking and constitutive recycling of AQP2 in stable cells expressing single and double mutations of AQP2 at S256 and S261. Cells expressing AQP2-S261A, S261D, S256A/S261A, S256A/S261D, S256D/S261A, and S256D/S261D were treated with VP (10 nM) and methyl-β-cyclodextrin (mβ-CD; 10 mM) for 20 min. Under unstimulated conditions, S256D/S261A and S256D/S261D accumulated on the plasma membrane (E and F) while AQP2-S261A, S261D, S256A/S261A, and S256A/S261D were located in the perinuclear cytoplasm (A–D). Upon stimulation with VP, S261A and S261D accumulated on the plasma membrane (G and H), similar to wild-type AQP2, while S256A/S261A and S256A/S261D remained in intracellular vesicles (I and J). S256D/S261A and S256D/S261D were located on the plasma membrane under all conditions (K and L). In all cell lines, a marked membrane accumulation was induced by treatment with methyl-β-cyclodextrin (from M–R). Results shown are representative of at least 3 independent experiments for each transfected cell line. B and H are duplicated images of Fig 2, E and F, and are included here for easy comparison. Bars = 20 μm.