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. 2003 May;121(5):413–425. doi: 10.1085/jgp.200308803

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Copyright © 2003, The Rockefeller University Press

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Figure 10. — Aquaporin 2–containing endosomes from control rat kidney show Hg²⁺-inhibitable water flux that is unchanged by addition of Anx4/Ca²⁺. (A) Endosomes enriched in AQP2 were isolated from rat kidney after in vivo loading with carboxyfluorescein as described in materials and methods. Western blotting of equivalent protein loaded lanes (10 μg) revealed that the purified fraction was fivefold enriched for AQP2 (lane 2) when compared with homogenate (lane 1). (B) AQP2-containing endosomes exhibited rapid water transport which was completely inhibited by treatment of membranes with 1 mM HgCl₂ for 30 min (C) AQP2-containing endosomes treated with Ca²⁺ and Anx4 for 30 min exhibited water transport kinetics that were identical to untreated membranes. Anx4 was added to endosomes at 1:1 and 2:1 protein ratios (2:1 shown). Results shown are representative of three separate experiments.